Extracellular matrix and cell adhesion molecules

ABSTRACT

The invention provides human extracellular matrix and cell adhesion molecules (XMAD) and polynucleotides which identify and encode XMAD. The invention also provides expression vectors, host cells, antibodies, agonists, and antagonists. The invention also provides methods for diagnosing, treating, or preventing disorders associated with expression of XMAD.

TECHNICAL FIELD

[0001] This invention relates to nucleic acid and amino acid sequences of extracellular matrix and cell adhesion molecules and to the use of these sequences in the diagnosis, treatment, and prevention of genetic, autoimmune/inflammation, and cell proliferative disorders, including cancer, and in the assessment of the effects of exogenous compounds on the expression of nucleic acid and amino acid sequences of extracellular matrix and cell adhesion molecules.

BACKGROUND OF THE INVENTION

[0002] Extracellular Matrix Proteins

[0003] The extracellular matrix (ECM) is a complex network of glycoproteins, polysaccharides, proteoglycans, and other macromolecules that are secreted from the cell into the extracellular space. The ECM remains in close association with the cell surface and provides a supportive meshwork that profoundly influences cell shape, motility, strength, flexibility, and adhesion. In fact, adhesion of a cell to its surrounding matrix is required for cell survival except in the case of metastatic tumor cells, which have overcome the need for cell-ECM anchorage. This phenomenon suggests that the ECM plays a critical role in the molecular mechanisms of growth control and metastasis. (Reviewed in Ruoslahti, E. (1996) Sci. Am. 275:72-77.) Furthermore, the ECM determines the structure and physical properties of connective tissue and is particularly important for morphogenesis and other processes associated with embryonic development and pattern formation.

[0004] The collagens comprise a family of ECM proteins that provide structure to bone, teeth, skin, ligaments, tendons, cartilage, blood vessels, and basement membranes. Multiple collagen proteins have been identified. Three collagen molecules fold together in a triple helix stabilized by interchain disulfide bonds. Bundles of these triple helices then associate to form fibrils.

[0005] Elastin and related proteins confer elasticity to tissues such as skin, blood vessels, and lungs. Elastin is a highly hydrophobic protein of about 750 amino acids that is rich in proline and glycine residues. Elastin molecules are highly cross-linked, forming an extensive extracellular network of fibers and sheets. Elastin fibers are surrounded by a sheath of microfibrils which are composed of a number of glycoproteins, including fibrillin.

[0006] Fibronectin is a large ECM glycoprotein found in all vertebrates. Fibronectin exists as a dimer of two subunits, each containing about 2,500 amino acids. Each subunit folds into a rod-like structure containing multiple domains. The domains each contain multiple repeated modules, the most common of which is the type III fibronectin repeat. The type III fibronectin repeat is about 90 amino acids in length and is also found in other ECM proteins and in some plasma membrane and cytoplasmic proteins. Furthermore, some type III fibronectin repeats contain a characteristic tripeptide consisting of Arginine-Glycine-Aspartic acid (RGD). The RGD sequence is recognized by the integrin family of cell surface receptors and is also found in other ECM proteins. (Reviewed in Alberts, supra, pp. 986-987.)

[0007] Laminin is a major glycoprotein component of the basal lamina which underlies and supports epithelial cell sheets. Laminin is one of the first ECM proteins synthesized in the developing embryo. Laminin is an 850 kilodalton protein composed of three polypeptide chains joined in the shape of a cross by disulfide bonds. Laminin is especially important for angiogenesis and, in particular, for guiding the formation of capillaries. (Reviewed in Alberts, B., et al. (1994) Molecular Biology of the Cell, Garland Publishing, New York, N.Y., pp. 990-991.)

[0008] Many proteinaceous ECM components are proteoglycans. Proteoglycans are composed of unbranched polysaccharide chains (glycosaminoglycans) attached to protein cores. Common proteoglycans include aggrecan, betaglycan, decorin, perlecan, serglycin, and syndecan-1. Some of these molecules not only provide mechanical support, but also bind to extracellular signaling molecules, such as fibroblast growth factor and transforming growth factor β, suggesting a role for proteoglycans in cell-cell communication. (Reviewed in Alberts, supra, pp. 973-978.)

[0009] Dentin phosphoryn (DPP) is a major component of the dentin ECM. DPP is a proteoglycan that is synthesized and expressed by odontoblasts (Gu, K., et al. (1998) Eur. J. Oral Sci. 106:1043-1047). DPP is believed to nucleate or modulate the formation of hydroxyapatite crystals.

[0010] Mucins are highly glycosylated glycoproteins that are the major structural component of the mucus gel. The physiological functions of mucins are cytoprotection, mechanical protection, maintenance of viscosity in secretions, and cellular recognition. MUC6 is a human gastric mucin that is also found in gall bladder, pancreas, seminal vesicles, and female reproductive tract (Toribara, N. W., et al. (1997) J. Biol. Chem. 272:16398-16403). The MUC6 gene has been mapped to human chromosome 11 (Toribara, N. W., et al. (1993) J. Biol. Chem. 268:5879-5885). Hemomucin is a novel Drosophila surface mucin that may be involved in the induction of antibacterial effector molecules (Theopold, U., et al. (1996) J. Biol. Chem. 217:12708-12715).

[0011] Extracellular matrix proteins may regulate cellular protein activity in a variety of ways. Reversible protein phosphorylation is the primary method for regulating protein activity in eukaryotic cells. In general, proteins are activated by phosphorylation in response to extracellular signals such as hormones, neurotransmitters, and growth and differentiation factors. The activated proteins initiate the cell's intracellular response by way of intracellular signaling pathways and second messenger molecules such as cyclic nucleotides, calcium-calmodulin, inositol, and various mitogens, that regulate protein phosphorylation.

[0012] Adhesion-Associated Proteins

[0013] The surface of a cell is rich in transmembrane proteoglycans, glycoproteins, glycolipids, and receptors. These macromolecules mediate adhesion with other cells and with components of the ECM. The interaction of the cell with its surroundings profoundly influences cell shape, strength, flexibility, motility, and adhesion. These dynamic properties are intimately associated with signal transduction pathways controlling cell proliferation and differentiation, tissue construction, and embryonic development.

[0014] Cadherins comprise a family of calcium-dependent glycoproteins that function in mediating cell-cell adhesion in virtually all solid tissues of multicellular organisms. These proteins share multiple repeats of a cadherin-specific motif, and the repeats form the folding units of the cadherin ECM. Cadherin molecules cooperate to form focal contacts, or adhesion plaques, between adjacent epithelial cells. The cadherin family includes the classical cadherins and protocadherins. Classical cadherins include the E-cadherin, N-cadherin, and P-cadherin subfamilies. E-cadherin is present on many types of epithelial cells and is especially important for embryonic development. P-cadherin is present on cells of the placenta and epidermis. Recent studies report that protocadherins are involved in a variety of cell-cell interactions (Suzuki, S. T. (1996) J. Cell Sci. 109:2609-2611). The intracellular anchorage of cadherins is regulated by their dynamic association with catenins, a family of cytoplasmic signal transduction proteins associated with the actin cytoskeleton. The anchorage of cadherins to the actin cytoskeleton appears to be regulated by protein tyrosine phosphorylation, and the cadherins are the target of phosphorylation-induced junctional disassembly (Aberle, H., et al. (1996) J. Cell. Biochem. 61:514-523).

[0015] Integrins are ubiquitous transmembrane adhesion molecules that link the ECM to the internal cytoskeleton. Integrins are composed of two noncovalently associated transmembrane glycoprotein subunits called α and β. Integrins function as receptors that play a role in signal transduction. For example, binding of integrin to its extracellular ligand may stimulate changes in intracellular calcium levels or protein kinase activity (Sjaastad, M. D. and Nelson, W. J. (1997) BioEssays 19:47-55).

[0016] Lectins comprise a ubiquitous family of extracellular glycoproteins which bind cell surface carbohydrates specifically and reversibly, resulting in the agglutination of cells. (Reviewed in Drickamer, K. and Taylor, M. E. (1993) Annu. Rev. Cell Biol. 9:237-264.) This function is particularly important for activation of the immune response. Lectins mediate the agglutination and mitogenic stimulation of lymphocytes at sites of inflammation (Lasky, L. A. (1991) J. Cell. Biochem. 45:139-146; Paietta, E., et al. (1989) J. Immunol. 143:2850-2857). C-type lectin domains are found in a variety of proteins, including selectins and lecticans. Lecticans are a family of chondroitin sulfate proteoglycans that include aggrecan, versican, neurocan, and brevican. All C-type lectin proteins are involved in protein-protein interactions (Aspberg, A., et al. (1997) Proc. Natl. Acad. Sci. USA 94:10116-10121). A novel macrophage-restricted C-type lectin protein has been cloned from mouse tissue. It is a type II transmembrane protein with one extracellular C-type lectin domain (Balch, S. G., et al. (1998) J. Biol. Chem. 273:18656-18664).

[0017] Toposome is a cell-adhesion glycoprotein isolated from mesenchyme-blastula embryos. Toposome precursors including vitellogenin promote cell adhesion of dissociated blastula cells.

[0018] LRRs are sequence motifs, approximately 22-28 amino acids in length, found in proteins with a large variety of functions and cellular locations. Proteins containing LRRs are all thought to be involved in protein-protein interactions. The crystal structure of LRRs has been studied and found to correspond to beta-alpha structural units. These structural units form a parallel beta sheet with one surface exposed to solvent. In this way an LRR-containing protein acquires a nonglobular shape (Kobe, B. and Deisenhofer, J. (1994) Trends Biochem. Sci. 19:415-421). There is evidence to suggest LRRs function in signal transduction and cellular adhesion as well as in protein-protein interactions (Gay, N. J., et al. (1991) FEBS Lett. 29:87-91).

[0019] Various proteins such as those encoded by the Drosophila armadillo gene and the human APC gene contain amino acid repeats that interact with β-catenins. The armadillo gene is required for pattern formation within the embryonic segments and imaginal discs and is highly conserved. It is 63% identical to a human protein, plakoglobin, which is involved in adhesive junctions joining epithelial and other cells (Peifer, M. and Wieschaus, E. (1990) Cell 63:1167-1176). APC gene mutations appear to initiate inherited forms of human colorectal cancer and sporadic forms of colorectal and gastric cancer (Rubinfeld, B., et al. (1993) Science 262:1731-1734). The fact that the protein encoded by APC interacts with catenin suggests a link between tumor initiation and cell adhesion (Su, L. K., et al. (1993) Science 262:1734-1737).

[0020] SH3 is a 60-70 amino acid motif found in a variety of signal transduction and cytoskeletal proteins. The SH3 domain is involved in mediating protein-protein interactions. Evidence suggests that the SH3 domains recognize a family of related domains or proteins in a variety of different tissues and species. One novel SH3 domain-containing protein is the 52 kilodalton focal adhesion protein (FAP52 or p52). FAP52 is localized to focal adhesions, specialized membrane domains in cultured cells that mediate the attachment of cells to the growth substratum and ECM. Focal adhesions consist of structural proteins, integrins, regulatory molecules, and signaling molecules and are involved in cell signaling. FAP52 may form part of this multimolecular complex that comprises focal adhesion sites (Merilainent, J., et al. (1997) J. Biol. Chem. 272:23278-23284).

[0021] The discovery of new extracellular matrix and cell adhesion molecules and the polynucleotides encoding them satisfies a need in the art by providing new compositions which are useful in the diagnosis, prevention, and treatment of genetic, autoimmune/inflammation, and cell proliferative disorders, including cancer, and in the assessment of the effects of exogenous compounds on the expression of nucleic acid and amino acid sequences of extracellular matrix and cell adhesion molecules.

SUMMARY OF THE INVENTION

[0022] The invention features purified polypeptides, extracellular matrix and cell adhesion molecules, referred to collectively as “XMAD” and individually as “XMAD-1,” “XMAD-2,” “XMAD-3,” “XMAD-4,” “XMAD-5,” “XMAD-6,” “XMAD-7,” “XMAD-8,” “XMAD-9,” “XMAD-10,” “XMAD-11,” “XMAD-12,” “XMAD-13,” “XMAD-14,” “XMAD-15,” “XMAD-16,” “XMAD-17,” “XMAD-18,” “XMAD-19,” “XMAD-20,” and “XMAD-21.” In one aspect, the invention provides an isolated polypeptide comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21. In one alternative, the invention provides an isolated polypeptide comprising the amino acid sequence of SEQ ID NO:1-21.

[0023] The invention further provides an isolated polynucleotide encoding a polypeptide comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21. In one alternative, the polynucleotide encodes a polypeptide selected from the group consisting of SEQ ID NO:1-21. In another alternative, the polynucleotide is selected from the group consisting of SEQ ID NO:22-42.

[0024] Additionally, the invention provides a recombinant polynucleotide comprising a promoter sequence operably linked to a polynucleotide encoding a polypeptide comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21. In one alternative, the invention provides a cell transformed with the recombinant polynucleotide. In another alternative, the invention provides a transgenic organism comprising the recombinant polynucleotide.

[0025] The invention also provides a method for producing a polypeptide comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21. The method comprises a) culturing a cell under conditions suitable for expression of the polypeptide, wherein said cell is transformed with a recombinant polynucleotide comprising a promoter sequence operably linked to a polynucleotide encoding the polypeptide, and b) recovering the polypeptide so expressed.

[0026] Additionally, the invention provides an isolated antibody which specifically binds to a polypeptide comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21.

[0027] The invention further provides an isolated polynucleotide comprising a polynucleotide sequence selected from the group consisting of a) a polynucleotide sequence selected from the group consisting of SEQ ID NO:22-42, b) a naturally occurring polynucleotide sequence having at least 90% sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID NO:22-42, c) a polynucleotide sequence complementary to a), d) a polynucleotide sequence complementary to b), and e) an RNA equivalent of a)-d). In one alternative, the polynucleotide comprises at least 60 contiguous nucleotides.

[0028] Additionally, the invention provides a method for detecting a target polynucleotide in a sample, said target polynucleotide having a sequence of a polynucleotide comprising a polynucleotide sequence selected from the group consisting of a) a polynucleotide sequence selected from the group consisting of SEQ ID NO:22-42, b) a naturally occurring polynucleotide sequence having at least 90% sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID NO:22-42, c) a polynucleotide sequence complementary to a), d) a polynucleotide sequence complementary to b), and e) an RNA equivalent of a)-d). The method comprises a) hybridizing the sample with a probe comprising at least 20 contiguous nucleotides comprising a sequence complementary to said target polynucleotide in the sample, and which probe specifically hybridizes to said target polynucleotide, under conditions whereby a hybridization complex is formed between said probe and said target polynucleotide or fragments thereof, and b) detecting the presence or absence of said hybridization complex, and optionally, if present, the amount thereof. In one alternative, the probe comprises at least 60 contiguous nucleotides.

[0029] The invention further provides a method for detecting a target polynucleotide in a sample, said target polynucleotide having a sequence of a polynucleotide comprising a polynucleotide sequence selected from the group consisting of a) a polynucleotide sequence selected from the group consisting of SEQ ID NO:22-42, b) a naturally occurring polynucleotide sequence having at least 90% sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID NO:22-42, c) a polynucleotide sequence complementary to a), d) a polynucleotide sequence complementary to b), and e) an RNA equivalent of a)-d). The method comprises a) amplifying said target polynucleotide or fragment thereof using polymerase chain reaction amplification, and b) detecting the presence or absence of said amplified target polynucleotide or fragment thereof, and, optionally, if present, the amount thereof.

[0030] The invention further provides a composition comprising an effective amount of a polypeptide comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, and a pharmaceutically acceptable excipient In one embodiment, the composition comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1-21. The invention additionally provides a method of treating a disease or condition associated with decreased expression of functional XMAD, comprising administering to a patient in need of such treatment the composition.

[0031] The invention also provides a method for screening a compound for effectiveness as an agonist of a polypeptide comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21. The method comprises a) exposing a sample comprising the polypeptide to a compound, and b) detecting agonist activity in the sample. In one alternative, the invention provides a composition comprising an agonist compound identified by the method and a pharmaceutically acceptable excipient. In another alternative, the invention provides a method of treating a disease or condition associated with decreased expression of functional XMAD, comprising administering to a patient in need of such treatment the composition.

[0032] Additionally, the invention provides a method for screening a compound for effectiveness as an antagonist of a polypeptide comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21. The method comprises a) exposing a sample comprising the polypeptide to a compound, and b) detecting antagonist activity in the sample. In one alternative, the invention provides a composition comprising an antagonist compound identified by the method and a pharmaceutically acceptable excipient. In another alternative, the invention provides a method of treating a disease or condition associated with overexpression of functional XMAD, comprising administering to a patient in need of such treatment the composition.

[0033] The invention further provides a method of screening for a compound that specifically binds to a polypeptide comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21. The method comprises a) combining the polypeptide with at least one test compound under suitable conditions, and b) detecting binding of the polypeptide to the test compound, thereby identifying a compound that specifically binds to the polypeptide.

[0034] The invention further provides a method of screening for a compound that modulates the activity of a polypeptide comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21. The method comprises a) combining the polypeptide with at least one test compound under conditions permissive for the activity of the polypeptide, b) assessing the activity of the polypeptide in the presence of the test compound, and c) comparing the activity of the polypeptide in the presence of the test compound with the activity of the polypeptide in the absence of the test compound, wherein a change in the activity of the polypeptide in the presence of the test compound is indicative of a compound that modulates the activity of the polypeptide.

[0035] The invention further provides a method for screening a compound for effectiveness in altering expression of a target polynucleotide, wherein said target polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO:22-42, the method comprising a) exposing a sample comprising the target polynucleotide to a compound, and b) detecting altered expression of the target polynucleotide.

[0036] The invention further provides a method for assessing toxicity of a test compound, said method comprising a) treating a biological sample containing nucleic acids with the test compound; b) hybridizing the nucleic acids of the treated biological sample with a probe comprising at least 20 contiguous nucleotides of a polynucleotide comprising a polynucleotide sequence selected from the group consisting of i) a polynucleotide sequence selected from the group consisting of SEQ ID NO:22-42, ii) a naturally occurring polynucleotide sequence having at least 90% sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID NO:22-42, iii) a polynucleotide sequence complementary to i), iv) a polynucleotide sequence complementary to ii), and v) an RNA equivalent of i)-iv). Hybridization occurs under conditions whereby a specific hybridization complex is formed between said probe and a target polynucleotide in the biological sample, said target polynucleotide comprising a polynucleotide sequence selected from the group consisting of i) a polynucleotide sequence selected from the group consisting of SEQ ID NO:22-42, ii) a naturally occurring polynucleotide sequence having at least 90% sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID NO:22-42, iii) a polynucleotide sequence complementary to i), iv) a polynucleotide sequence complementary to ii), and v) an RNA equivalent of i)-iv). Alternatively, the target polynucleotide comprises a fragment of a polynucleotide sequence selected from the group consisting of i)-v) above; c) quantifying the amount of hybridization complex; and d) comparing the amount of hybridization complex in the treated biological sample with the amount of hybridization complex in an untreated biological sample, wherein a difference in the amount of hybridization complex in the treated biological sample is indicative of toxicity of the test compound.

BRIEF DESCRIPTION OF THE TABLES

[0037] Table 1 shows polypeptide and nucleotide sequence identification numbers (SEQ ID NOs), clone identification numbers (clone IDs), cDNA libraries, and cDNA fragments used to assemble full-length sequences encoding XMAD.

[0038] Table 2 shows features of each polypeptide sequence, including potential motifs, homologous sequences, and methods, algorithms, and searchable databases used for analysis of XMAD.

[0039] Table 3 shows selected fragments of each nucleic acid sequence; the tissue-specific expression patterns of each nucleic acid sequence as determined by northern analysis; diseases, disorders, or conditions associated with these tissues; and the vector into which each cDNA was cloned.

[0040] Table 4 describes the tissues used to construct the cDNA libraries from which cDNA clones encoding XMAD were isolated.

[0041] Table 5 shows the tools, programs, and algorithms used to analyze the polynucleotides and polypeptides of the invention, along with applicable descriptions, references, and threshold parameters.

DESCRIPTION OF THE INVENTION

[0042] Before the present proteins, nucleotide sequences, and methods are described, it is understood that this invention is not limited to the particular machines, materials and methods described, as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention which will be limited only by the appended claims.

[0043] It must be noted that as used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural reference unless the context clearly dictates otherwise. Thus, for example, a reference to “a host cell” includes a plurality of such host cells, and a reference to “an antibody” is a reference to one or more antibodies and equivalents thereof known to those skilled in the art, and so forth.

[0044] Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any machines, materials, and methods similar or equivalent to those described herein can be used to practice or test the present invention, the preferred machines, materials and methods are now described. All publications mentioned herein are cited for the purpose of describing and disclosing the cell lines, protocols, reagents and vectors which are reported in the publications and which might be used in connection with the invention. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.

[0045] Definitions “XMAD” refers to the amino acid sequences of substantially purified XMAD obtained from any species, particularly a mammalian species, including bovine, ovine, porcine, murine, equine, and human, and from any source, whether natural, synthetic, semi-synthetic, or recombinant.

[0046] The term “agonist” refers to a molecule which intensifies or mimics the biological activity of XMAD. Agonists may include proteins, nucleic acids, carbohydrates, small molecules, or any other compound or composition which modulates the activity of XMAD either by directly interacting with XMAD or by acting on components of the biological pathway in which XMAD participates.

[0047] An “allelic variant” is an alternative form of the gene encoding XMAD. Allelic variants may result from at least one mutation in the nucleic acid sequence and may result in altered mRNAs or in polypeptides whose structure or function may or may not be altered. A gene may have none, one, or many allelic variants of its naturally occurring form. Common mutational changes which give rise to allelic variants are generally ascribed to natural deletions, additions, or substitutions of nucleotides. Each of these types of changes may occur alone, or in combination with the others, one or more times in a given sequence.

[0048] “Altered” nucleic acid sequences encoding XMAD include those sequences with deletions, insertions, or substitutions of different nucleotides, resulting in a polypeptide the same as XMAD or a polypeptide with at least one functional characteristic of XMAD. Included within this definition are polymorphisms which may or may not be readily detectable using a particular oligonucleotide probe of the polynucleotide encoding XMAD, and improper or unexpected hybridization to allelic variants, with a locus other than the normal chromosomal locus for the polynucleotide sequence encoding XMAD. The encoded protein may also be “altered,” and may contain deletions, insertions, or substitutions of amino acid residues which produce a silent change and result in a functionally equivalent XMAD. Deliberate amino acid substitutions may be made on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity, and/or the amphipathic nature of the residues, as long as the biological or immunological activity of XMAD is retained. For example, negatively charged amino acids may include aspartic acid and glutamic acid, and positively charged amino acids may include lysine and arginine. Amino acids with uncharged polar side chains having similar hydrophilicity values may include: asparagine and glutamine; and serine and threonine. Amino acids with uncharged side chains having similar hydrophilicity values may include: leucine, isoleucine, and valine; glycine and alanine; and phenylalanine and tyrosine.

[0049] The terms “amino acid” and “amino acid sequence” refer to an oligopeptide, peptide, polypeptide, or protein sequence, or a fragment of any of these, and to naturally occurring or synthetic molecules. Where “amino acid sequence” is recited to refer to a sequence of a naturally occurring protein molecule, “amino acid sequence” and like terms are not meant to limit the amino acid sequence to the complete native amino acid sequence associated with the recited protein molecule.

[0050] “Amplification” relates to the production of additional copies of a nucleic acid sequence. Amplification is generally carried out using polymerase chain reaction (PCR) technologies well known in the art.

[0051] The term “antagonist” refers to a molecule which inhibits or attenuates the biological activity of XMAD. Antagonists may include proteins such as antibodies, nucleic acids, carbohydrates, small molecules, or any other compound or composition which modulates the activity of XMAD either by directly interacting with XMAD or by acting on components of the biological pathway in which XMAD participates.

[0052] The term “antibody” refers to intact immunoglobulin molecules as well as to fragments thereof, such as Fab, F(ab′)₂, and Fv fragments, which are capable of binding an epitopic determinant. Antibodies that bind XMAD polypeptides can be prepared using intact polypeptides or using fragments containing small peptides of interest as the immunizing antigen. The polypeptide or oligopeptide used to immunize an animal (e.g., a mouse, a rat, or a rabbit) can be derived from the translation of RNA, or synthesized chemically, and can be conjugated to a carrier protein if desired. Commonly used carriers that arc chemically coupled to peptides include bovine serum albumin, thyroglobulin, and keyhole limpel hemocyanin (KLH). The coupled peptide is then used to immunize the animal.

[0053] The term “antigenic determinant” refers to that region of a molecule (i.e., an epitope) that makes contact with a particular antibody. When a protein or a fragment of a protein is used to immunize a host animal, numerous regions of the protein may induce the production of antibodies which bind specifically to antigenic determinants (particular regions or three-dimensional structures on the protein). An antigenic determinant may compete with the intact antigen (i.e., the immunogen used to elicit the immune response) for binding to an antibody.

[0054] The term “antisense” refers to any composition capable of base-pairing with the “sense” (coding) strand of a specific nucleic acid sequence. Antisense compositions may include DNA; RNA; peptide nucleic acid (PNA); oligonucleotides having modified backbone linkages such as phosphorothioates, methylphosphonates, or benzylphosphonates; oligonucleotides having modified sugar groups such as 2′-methoxyethyl sugars or 2′-methoxyethoxy sugars; or oligonucleotides having modified bases such as 5-methyl cytosine, 2′-deoxyuracil, or 7-deaza-2′-deoxyguanosine. Antisense molecules may be produced by any method including chemical synthesis or transcription. Once introduced into a cell, the complementary antisense molecule base-pairs with a naturally occurring nucleic acid sequence produced by the cell to form duplexes which block either transcription or translation. The designation “negative” or “minus” can refer to the antisense strand, and the designation “positive” or “plus” can refer to the sense strand of a reference DNA molecule.

[0055] The term “biologically active” refers to a protein having structural, regulatory, or biochemical functions of a naturally occurring molecule. Likewise, “immunologically active” or “immunogenic” refers to the capability of the natural, recombinant, or synthetic XMAD, or of any oligopeptide thereof, to induce a specific immune response in appropriate animals or cells and to bind with specific antibodies.

[0056] “Complementary” describes the relationship between two single-stranded nucleic acid sequences that anneal by base-pairing. For example, 5′-AGT-3′ pairs with its complement, 3′-TCA-5′.

[0057] A “composition comprising a given polynucleotide sequence” and a “composition comprising a given amino acid sequence” refer broadly to any composition containing the given polynucleotide or amino acid sequence. The composition may comprise a dry formulation or an aqueous solution. Compositions comprising polynucleotide sequences encoding XMAD or fragments of XMAD may be employed as hybridization probes. The probes may be stored in freeze-dried form and may be associated with a stabilizing agent such as a carbohydrate. In hybridizations, the probe may be deployed in an aqueous solution containing salts (e.g., NaCl), detergents (e.g., sodium dodecyl sulfate; SDS), and other components (e.g., Denhardt's solution, dry milk, salmon sperm DNA, etc.).

[0058] “Consensus sequence” refers to a nucleic acid sequence which has been subjected to repeated DNA sequence analysis to resolve uncalled bases, extended using the XL-PCR kit (Applied Biosystems, Foster City Calif.) in the 5′ and/or the 3′ direction, and resequenced, or which has been assembled from one or more overlapping cDNA, EST, or genomic DNA fragments using a computer program for fragment assembly, such as the GELVIEW fragment assembly system (GCG, Madison Wis.) or Phrap (University of Washington, Seattle Wash.). Some sequences have been both extended and assembled to produce the consensus sequence.

[0059] “Conservative amino acid substitutions” are those substitutions that are predicted to least interfere with the properties of the original protein, i.e., the structure and especially the function of the protein is conserved and not significantly changed by such substitutions. The table below shows amino acids which may be substituted for an original amino acid in a protein and which are regarded as conservative amino acid substitutions. Original Residue Conservative Substitution Ala Gly, Ser Arg His, Lys Asn Asp, Gln, His Asp Asn, Glu Cys Ala, Ser Gln Asn, Glu, His Glu Asp, Gln, His Gly Ala His Asn, Arg, Gln, Glu Ile Leu, Val Leu Ile, Val Lys Arg, Gln, Glu Met Leu, Ile Phe His, Met, Leu, Trp, Tyr Ser Cys, Thr Thr Ser, Val Trp Phe, Tyr Tyr His, Phe, Trp Val Ile, Leu, Thr

[0060] Conservative amino acid substitutions generally maintain (a) the structure of the polypeptide backbone in the area of the substitution, for example, as a beta sheet or alpha helical conformation, (b) the charge or hydrophobicity of the molecule at the site of the substitution, and/or (c) the bulk of the side chain.

[0061] A “deletion” refers to a change in the amino acid or nucleotide sequence that results in the absence of one or more amino acid residues or nucleotides.

[0062] The term “derivative” refers to a chemically modified polynucleotide or polypeptide. Chemical modifications of a polynucleotide sequence can include, for example, replacement of hydrogen by an alkyl, acyl, hydroxyl, or amino group. A derivative polynucleotide encodes a polypeptide which retains at least one biological or immunological function of the natural molecule. A derivative polypeptide is one modified by glycosylation, pegylation, or any similar process that retains at least one biological or immunological function of the polypeptide from which it was derived.

[0063] A “detectable label” refers to a reporter molecule or enzyme that is capable of generating a measurable signal and is covalently or noncovalently joined to a polynucleotide or polypeptide.

[0064] A “fragment” is a unique portion of XMAD or the polynucleotide encoding XMAD which is identical in sequence to but shorter in length than the parent sequence. A fragment may comprise up to the entire length of the defined sequence, minus one nucleotide/amino acid residue. For example, a fragment may comprise from 5 to 1000 contiguous nucleotides or amino acid residues. A fragment used as a probe, primer, antigen, therapeutic molecule, or for other purposes, may be at least 5, 10, 15, 16, 20, 25, 30, 40, 50, 60, 75, 100, 150, 250 or at least 500 contiguous nucleotides or amino acid residues in length. Fragments may be preferentially selected from certain regions of a molecule. For example, a polypeptide fragment may comprise a certain length of contiguous amino acids selected from the first 250 or 500 amino acids (or first 25% or 50% of a polypeptide) as shown in a certain defined sequence. Clearly these lengths are exemplary, and any length that is supported by the specification, including the Sequence Listing, tables, and figures, may be encompassed by the present embodiments.

[0065] A fragment of SEQ ID NO:22-42 comprises a region of unique polynucleotide sequence that specifically identifies SEQ ID NO:22-42, for example, as distinct from any other sequence in the genome from which the fragment was obtained. A fragment of SEQ ID NO:22-42 is useful, for example, in hybridization and amplification technologies and in analogous methods that distinguish SEQ ID NO:22-42 from related polynucleotide sequences. The precise length of a fragment of SEQ ID NO:22-42 and the region of SEQ ID NO:22-42 to which the fragment corresponds are routinely determinable by one of ordinary skill in the art based on the intended purpose for the fragment.

[0066] A fragment of SEQ ID NO:1-21 is encoded by a fragment of SEQ ID NO:22-42. A fragment of SEQ ID NO:1-21 comprises a region of unique amino acid sequence that specifically identifies SEQ ID NO:1-21. For example, a fragment of SEQ ID NO:1-21 is useful as an immunogenic peptide for the development of antibodies that specifically recognize SEQ ID NO:1-21. The precise length of a fragment of SEQ ID NO:1-21 and the region of SEQ ID NO:1-21 to which the fragment corresponds are routinely determinable by one of ordinary skill in the art based on the intended purpose for the fragment.

[0067] A “full-length” polynucleotide sequence is one containing at least a translation initiation codon (e.g., methionine) followed by an open reading frame and a translation termination codon. A “full-length” polynucleotide sequence encodes a “full-length” polypeptide sequence.

[0068] “Homology” refers to sequence similarity or, interchangeably, sequence identity, between two or more polynucleotide sequences or two or more polypeptide sequences.

[0069] The terms “percent identity” and “% identity,” as applied to polynucleotide sequences, refer to the percentage of residue matches between at least two polynucleotide sequences aligned using a standardized algorithm. Such an algorithm may insert, in a standardized and reproducible way, gaps in the sequences being compared in order to optimize alignment between two sequences, and therefore achieve a more meaningful comparison of the two sequences.

[0070] Percent identity between polynucleotide sequences may be determined using the default parameters of the CLUSTAL V algorithm as incorporated into the MEGALIGN version 3.12e sequence alignment program. This program is part of the LASERGENE software package, a suite of molecular biological analysis programs (DNASTAR, Madison Wis.). CLUSTAL V is described in Higgins, D. G. and P. M. Sharp (1989) CABIOS 5:151-153 and in Higgins, D. G. et al. (1992) CABIOS 8:189-191. For pairwise alignments of polynucleotide sequences, the default parameters are set as follows: Ktuple=2, gap penalty=5, window=4, and “diagonals saved”=4. The “weighted” residue weight table is selected as the default. Percent identity is reported by CLUSTAL V as the “percent similarity” between aligned polynucleotide sequences.

[0071] Alternatively, a suite of commonly used and freely available sequence comparison algorithms is provided by the National Center for Biotechnology Information (NCBI) Basic Local Alignment Search Tool (BLAST) (Altschul, S. F. et al. (1990) J. Mol. Biol. 215:403410), which is available from several sources, including the NCBI, Bethesda, Md., and on the Internet at http://www.ncbi.nlm.nih.gov/BLAST/. The BLAST software suite includes various sequence analysis programs including “blastn,” that is used to align a known polynucleotide sequence with other polynucleotide sequences from a variety of databases. Also available is a tool called “BLAST 2 Sequences” that is used for direct pairwise comparison of two nucleotide sequences. “BLAST 2 Sequences” can be accessed and used interactively at http://www.ncbi.nlm.nih.gov/gorf/bl2.html. The “BLAST 2 Sequences” tool can be used for both blastn and blastp (discussed below). BLAST programs are commonly used with gap and other parameters set to default settings. For example, to compare two nucleotide sequences, one may use blastn with the “BLAST 2 Sequences” tool Version 2.0.12 (Apr. 21, 2000) set at default parameters. Such default parameters may be, for example:

[0072] Matrix: BLOSUM62

[0073] Reward for match: 1

[0074] Penalty for mismatch: −2

[0075] Open Gap: 5 and Extension Gap: 2 penalties

[0076] Gap x drop-off: 50

[0077] Expect: 10

[0078] Word Size: 11

[0079] Filter: on

[0080] Percent identity may be measured over the length of an entire defined sequence, for example, as defined by a particular SEQ ID number, or may be measured over a shorter length, for example, over the length of a fragment taken from a larger, defined sequence, for instance, a fragment of at least 20, at least 30, at least 40, at least 50, at least 70, at least 100, or at least 200 contiguous nucleotides. Such lengths are exemplary only, and it is understood that any fragment length supported by the sequences shown herein, in the tables, figures, or Sequence Listing, may be used to describe a length over which percentage identity may be measured.

[0081] Nucleic acid sequences that do not show a high degree of identity may nevertheless encode similar amino acid sequences due to the degeneracy of the genetic code. It is understood that changes in a nucleic acid sequence can be made using this degeneracy to produce multiple nucleic acid sequences that all encode substantially the same protein.

[0082] The phrases “percent identity” and “% identity,” as applied to polypeptide sequences, refer to the percentage of residue matches between at least two polypeptide sequences aligned using a standardized algorithm. Methods of polypeptide sequence alignment are well-known. Some alignment methods take into account conservative amino acid substitutions. Such conservative substitutions, explained in more detail above, generally preserve the charge and hydrophobicity at the site of substitution, thus preserving the structure (and therefore function) of the polypeptide.

[0083] Percent identity between polypeptide sequences may be determined using the default parameters of the CLUSTAL V algorithm as incorporated into the MEGALIGN version 3.12e sequence alignment program (described and referenced above). For pairwise alignments of polypeptide sequences using CLUSTAL V, the default parameters are set as follows: Ktuple=1, gap penalty=3, window=5, and “diagonals saved”=5. The PAM250 matrix is selected as the default residue weight table. As with polynucleotide alignments, the percent identity is reported by CLUSTAL V as the “percent similarity” between aligned polypeptide sequence pairs.

[0084] Alternatively the NCBI BLAST software suite may be used. For example, for a pairwise comparison of two polypeptide sequences, one may use the “BLAST 2 Sequences” tool Version 2.0.12 (Apr. 21, 2000) with blastp set at default parameters. Such default parameters may be, for example:

[0085] Matrix: BLOSUM62

[0086] Open Gap: 11 and Extension Gap: 1 penalties

[0087] Gap x drop-off: 50

[0088] Expect: 10

[0089] Word Size: 3

[0090] Filler: on

[0091] Percent identity may be measured over the length of an entire defined polypeptide sequence, for example, as defined by a particular SEQ ID number, or may be measured over a shorter length, for example, over the length of a fragment taken from a larger, defined polypeptide sequence, for instance, a fragment of at least 15, at least 20, at least 30, at least 40, at least 50, at least 70 or at least 150 contiguous residues. Such lengths are exemplary only, and it is understood that any fragment length supported by the sequences shown herein, in the tables, figures or Sequence Listing, may be used to describe a length over which percentage identity may be measured.

[0092] “Human artificial chromosomes” (HACs) are linear microchromosomes which may contain DNA sequences of about 6 kb to 10 Mb in size, and which contain all of the elements required for chromosome replication, segregation and maintenance.

[0093] The term “humanized antibody” refers to an antibody molecule in which the amino acid sequence in the non-antigen binding regions has been altered so that the antibody more closely resembles a human antibody, and still retains its original binding ability.

[0094] “Hybridization” refers to the process by which a polynucleotide strand anneals with a complementary strand through base pairing under defined hybridization conditions. Specific hybridization is an indication that two nucleic acid sequences share a high degree of complementarity. Specific hybridization complexes form under permissive annealing conditions and remain hybridized after the “washing” step(s). The washing step(s) is particularly important in determining the stringency of the hybridization process, with more stringent conditions allowing less non-specific binding, i.e., binding between pairs of nucleic acid strands that are not perfectly matched. Permissive conditions for annealing of nucleic acid sequences are routinely determinable by one of ordinary skill in the art and may be consistent among hybridization experiments, whereas wash conditions may be varied among experiments to achieve the desired stringency, and therefore hybridization specificity. Permissive annealing conditions occur, for example, at 68° C. in the presence of about 6× SSC, about 1% (w/v) SDS, and about 100 μg/ml sheared, denatured salmon sperm DNA.

[0095] Generally, stringency of hybridization is expressed, in part, with reference to the temperature under which the wash step is carried out. Such wash temperatures are typically selected to be about 5° C. to 20° C. lower than the thermal melting point (T_(m)) for the specific sequence at a defined ionic strength and pH. The T_(m) is the temperature (under defined ionic strength and pH) at which 50% of the target sequence hybridizes to a perfectly matched probe. An equation for calculating T_(m) and conditions for nucleic acid hybridization are well known and can be found in Sambrook, J. et al., 1989, Molecular Cloning: A Laboratory Manual, 2^(nd) ed., vol. 1-3, Cold Spring Harbor Press, Plainview N.Y.; specifically see volume 2, chapter 9.

[0096] High stringency conditions for hybridization between polynucleotides of the present invention include wash conditions of 68° C. in the presence of about 0.2× SSC and about 0.1% SDS, for 1 hour. Alternatively, temperatures of about 65° C., 60° C., 55° C., or 42° C. may be used. SSC concentration may be varied from about 0.1 to 2× SSC, with SDS being present at about 0.1%. Typically, blocking reagents are used to block non-specific hybridization. Such blocking reagents include, for instance, sheared and denatured salmon sperm DNA at about 100-200 μg/ml. Organic solvent, such as formamide at a concentration of about 35-50% v/v, may also be used under particular circumstances, such as for RNA:DNA hybridizations. Useful variations on these wash conditions will be readily apparent to those of ordinary skill in the art. Hybridization, particularly under high stringency conditions, may be suggestive of evolutionary similarity between the nucleotides. Such similarity is strongly indicative of a similar role for the nucleotides and their encoded polypeptides.

[0097] The term “hybridization complex” refers to a complex formed between two nucleic acid sequences by virtue of the formation of hydrogen bonds between complementary bases. A hybridization complex may be formed in solution (e.g., C₀t or R₀t analysis) or formed between one nucleic acid sequence present in solution and another nucleic acid sequence immobilized on a solid support (e.g., paper, membranes, filters, chips, pins or glass slides, or any other appropriate substrate to which cells or their nucleic acids have been fixed).

[0098] The words “insertion” and “addition” refer to changes in an amino acid or nucleotide sequence resulting in the addition of one or more amino acid residues or nucleotides, respectively.

[0099] “Immune response” can refer to conditions associated with inflammation, trauma, immune disorders, or infectious or genetic disease, etc. These conditions can be characterized by expression of various factors, e.g., cytokines, chemokines, and other signaling molecules, which may affect cellular and systemic defense systems.

[0100] An “immunogenic fragment” is a polypeptide or oligopeptide fragment of XMAD which is capable of eliciting an immune response when introduced into a living organism, for example, a mammal. The term “immunogenic fragment” also includes any polypeptide or oligopeptide fragment of XMAD which is useful in any of the antibody production methods disclosed herein or known in the art.

[0101] The term “microarray” refers to an arrangement of a plurality of polynucleotides, polypeptides, or other chemical compounds on a substrate.

[0102] The terms “element” and “array element” refer to a polynucleotide, polypeptide, or other chemical compound having a unique and defined position on a microarray.

[0103] The term “modulate” refers to a change in the activity of XMAD. For example, modulation may cause an increase or a decrease in protein activity, binding characteristics, or any other biological, functional, or immunological properties of XMAD.

[0104] The phrases “nucleic acid” and “nucleic acid sequence” refer to a nucleotide, oligonucleotide, polynucleotide, or any fragment thereof. These phrases also refer to DNA or RNA of genomic or synthetic origin which may be single-stranded or double-stranded and may represent the sense or the antisense strand, to peptide nucleic acid (PNA), or to any DNA-like or RNA-like material.

[0105] “Operably linked” refers to the situation in which a first nucleic acid sequence is placed in a functional relationship with a second nucleic acid sequence. For instance, a promoter is operably linked to a coding sequence if the promoter affects the transcription or expression of the coding sequence. Operably linked DNA sequences may be in close proximity or contiguous and, where necessary to join two protein coding regions, in the same reading frame.

[0106] “Peptide nucleic acid” (PNA) refers to an antisense molecule or anti-gene agent which comprises an oligonucleotide of at least about 5 nucleotides in length linked to a peptide backbone of amino acid residues ending in lysine. The terminal lysine confers solubility to the composition. PNAs preferentially bind complementary single stranded DNA or RNA and stop transcript elongation, and may be pegylated to extend their lifespan in the cell.

[0107] “Post-translational modification” of an XMAD may involve lipidation, glycosylation, phosphorylation, acetylation, racemization, proteolytic cleavage, and other modifications known in the art. These processes may occur synthetically or biochemically. Biochemical modifications will vary by cell type depending on the enzymatic milieu of XMAD.

[0108] “Probe” refers to nucleic acid sequences encoding XMAD, their complements, or fragments thereof, which are used to detect identical, allelic or related nucleic acid sequences. Probes are isolated oligonucleotides or polynucleotides attached to a detectable label or reporter molecule. Typical labels include radioactive isotopes, ligands, chemiluminescent agents, and enzymes. “Primers” are short nucleic acids, usually DNA oligonucleotides, which may be annealed to a target polynucleotide by complementary base-pairing. The primer may then be extended along the target DNA strand by a DNA polymerase enzyme. Primer pairs can be used for amplification (and identification) of a nucleic acid sequence, e.g., by the polymerase chain reaction (PCR).

[0109] Probes and primers as used in the present invention typically comprise at least 15 contiguous nucleotides of a known sequence. In order to enhance specificity, longer probes and primers may also be employed, such as probes and primers that comprise at least 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, or at least 150 consecutive nucleotides of the disclosed nucleic acid sequences. Probes and primers may be considerably longer than these examples, and it is understood that any length supported by the specification, including the tables, figures, and Sequence Listing, may be used.

[0110] Methods for preparing and using probes and primers are described in the references, for example Sambrook, J. et al. (1989) Molecular Cloning: A Laboratory Manual, 2^(nd) ed., vol. 1-3, Cold Spring Harbor Press, Plainview N.Y.; Ausubel, F. M. et al. (1987) Current Protocols in Molecular Biology, Greene Publ. Assoc. & Wiley-Intersciences, New York N.Y.; Innis, M. et al. (1990) PCR Protocols, A Guide to Methods and Applications, Academic Press, San Diego Calif. PCR primer pairs can be derived from a known sequence, for example, by using computer programs intended for that purpose such as Primer (Version 0.5, 1991, Whitehead Institute for Biomedical Research, Cambridge Mass.).

[0111] Oligonucleotides for use as primers are selected using software known in the art for such purpose. For example, OLIGO 4.06 software is useful for the selection of PCR primer pairs of up to 100 nucleotides each, and for the analysis of oligonucleotides and larger polynucleotides of up to 5,000 nucleotides from an input polynucleotide sequence of up to 32 kilobases. Similar primer selection programs have incorporated additional features for expanded capabilities. For example, the PrimOU primer selection program (available to the public from the Genome Center at University of Texas South West Medical Center, Dallas Tex.) is capable of choosing specific primers from megabase sequences and is thus useful for designing primers on a genome-wide scope. The Primer3 primer selection program (available to the public from the Whitehead Institute/MIT Center for Genome Research, Cambridge Mass.) allows the user to input a “mispriming library,” in which sequences to avoid as primer binding sites are user-specified. Primer3 is useful, in particular, for the selection of oligonucleotides for microarrays. (The source code for the latter two primer selection programs may also be obtained from their respective sources and modified to meet the user's specific needs.) The PrimeGen program (available to the public from the UK Human Genome Mapping Project Resource Centre, Cambridge UK) designs primers based on multiple sequence alignments, thereby allowing selection of primers that hybridize to either the most conserved or least conserved regions of aligned nucleic acid sequences. Hence, this program is useful for identification of both unique and conserved oligonucleotides and polynucleotide fragments. The oligonucleotides and polynucleotide fragments identified by any of the above selection methods are useful in hybridization technologies, for example, as PCR or sequencing primers, microarray elements, or specific probes to identify fully or partially complementary polynucleotides in a sample of nucleic acids. Methods of oligonucleotide selection are not limited to those described above.

[0112] A “recombinant nucleic acid” is a sequence that is not naturally occurring or has a sequence that is made by an artificial combination of two or more otherwise separated segments of sequence. This artificial combination is often accomplished by chemical synthesis or, more commonly, by the artificial manipulation of isolated segments of nucleic acids, e.g., by genetic engineering techniques such as those described in Sambrook, supra. The term recombinant includes nucleic acids that have been altered solely by addition, substitution, or deletion of a portion of the nucleic acid. Frequently, a recombinant nucleic acid may include a nucleic acid sequence operably linked to a promoter sequence. Such a recombinant nucleic acid may be part of a vector that is used, for example, to transform a cell.

[0113] Alternatively, such recombinant nucleic acids may be part of a viral vector, e.g., based on a vaccinia virus, that could be use to vaccinate a mammal wherein the recombinant nucleic acid is expressed, inducing a protective immunological response in the mammal.

[0114] A “regulatory element” refers to a nucleic acid sequence usually derived from untranslated regions of a gene and includes enhancers, promoters, introns, and 5′ and 3′ untranslated regions (UTRs). Regulatory elements interact with host or viral proteins which control transcription, translation, or RNA stability.

[0115] “Reporter molecules” are chemical or biochemical moieties used for labeling a nucleic acid, amino acid, or antibody. Reporter molecules include radionuclides; enzymes; fluorescent, chemiluminescent, or chromogenic agents; substrates; cofactors; inhibitors; magnetic particles; and other moieties known in the art.

[0116] An “RNA equivalent,” in reference to a DNA sequence, is composed of the same linear sequence of nucleotides as the reference DNA sequence with the exception that all occurrences of the nitrogenous base thymine are replaced with uracil, and the sugar backbone is composed of ribose instead of deoxyribose.

[0117] The term “sample” is used in its broadest sense. A sample suspected of containing nucleic acids encoding XMAD, or fragments thereof, or XMAD itself, may comprise a bodily fluid; an extract from a cell, chromosome, organelle, or membrane isolated from a cell; a cell; genomic DNA, RNA, or cDNA, in solution or bound to a substrate; a tissue; a tissue print; etc.

[0118] The terms “specific binding” and “specifically binding” refer to that interaction between a protein or peptide and an agonist, an antibody, an antagonist, a small molecule, or any natural or synthetic binding composition. The interaction is dependent upon the presence of a particular structure of the protein, e.g., the antigenic determinant or epitope, recognized by the binding molecule. For example, if an antibody is specific for epitope “A,” the presence of a polypeptide comprising the epitope A, or the presence of free unlabeled A, in a reaction containing free labeled A and the antibody will reduce the amount of labeled A that binds to the antibody.

[0119] The term “substantially purified” refers to nucleic acid or amino acid sequences that are removed from their natural environment and are isolated or separated, and are at least 60% free, preferably at least 75% free, and most preferably at least 90% free from other components with which they are naturally associated.

[0120] A “substitution” refers to the replacement of one or more amino acid residues or nucleotides by different amino acid residues or nucleotides, respectively.

[0121] “Substrate” refers to any suitable rigid or semi-rigid support including membranes, filters, chips, slides, wafers, fibers, magnetic or nonmagnetic beads, gels, tubing, plates, polymers, microparticles and capillaries. The substrate can have a variety of surface forms, such as wells, trenches, pins, channels and pores, to which polynucleotides or polypeptides are bound.

[0122] A “transcript image” refers to the collective pattern of gene expression by a particular cell type or tissue under given conditions at a given time.

[0123] “Transformation” describes a process by which exogenous DNA is introduced into a recipient cell. Transformation may occur under natural or artificial conditions according to various methods well known in the art, and may rely on any known method for the insertion of foreign nucleic acid sequences into a prokaryotic or eukaryotic host cell. The method for transformation is selected based on the type of host cell being transformed and may include, but is not limited to, bacteriophage or viral infection, electroporation, heat shock, lipofection, and particle bombardment. The term “transformed” cells includes stably transformed cells in which the inserted DNA is capable of replication either as an autonomously replicating plasmid or as part of the host chromosome, as well as transiently transformed cells which express the inserted DNA or RNA for limited periods of time.

[0124] A “transgenic organism,” as used herein, is any organism, including but not limited to animals and plants, in which one or more of the cells of the organism contains heterologous nucleic acid introduced by way of human intervention, such as by transgenic techniques well known in the art. The nucleic acid is introduced into the cell, directly or indirectly by introduction into a precursor of the cell, by way of deliberate genetic manipulation, such as by microinjection or by infection with a recombinant virus. The term genetic manipulation does not include classical cross-breeding, or in vitro fertilization, but rather is directed to the introduction of a recombinant DNA molecule. The transgenic organisms contemplated in accordance with the present invention include bacteria, cyanobacteria, fungi, plants, and animals. The isolated DNA of the present invention can be introduced into the host by methods known in the art, for example infection, transfection, transformation or transconjugation. Techniques for transferring the DNA of the present invention into such organisms are widely known and provided in references such as Sambrook, J. et al. (1989), supra.

[0125] A “variant” of a particular nucleic acid sequence is defined as a nucleic acid sequence having at least 40% sequence identity to the particular nucleic acid sequence over a certain length of one of the nucleic acid sequences using blastn with the “BLAST 2 Sequences” tool Version 2.0.9 (May 7, 1999) set at default parameters. Such a pair of nucleic acids may show, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95% or at least 98% or greater sequence identity over a certain defined length. A variant may be described as, for example, an “allelic” (as defined above), “splice,” “species,” or “polymorphic” variant. A splice variant may have significant identity to a reference molecule, but will generally have a greater or lesser number of polynucleotides due to alternative splicing of exons during mRNA processing. The corresponding polypeptide may possess additional functional domains or lack domains that are present in the reference molecule. Species variants are polynucleotide sequences that vary from one species to another. The resulting polypeptides generally will have significant amino acid identity relative to each other. A polymorphic variant is a variation in the polynucleotide sequence of a particular gene between individuals of a given species. Polymorphic variants also may encompass “single nucleotide polymorphisms” (SNPs) in which the polynucleotide sequence varies by one nucleotide base. The presence of SNPs may be indicative of, for example, a certain population, a disease state, or a propensity for a disease state.

[0126] A “variant” of a particular polypeptide sequence is defined as a polypeptide sequence having at least 40% sequence identity to the particular polypeptide sequence over a certain length of one of the polypeptide sequences using blastp with the “BLAST 2 Sequences” tool Version 2.0.9 (May 7, 1999) set at default parameters. Such a pair of polypeptides may show, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 98% or greater sequence identity over a certain defined length of one of the polypeptides.

[0127] The Invention

[0128] The invention is based on the discovery of new human extracellular matrix and cell adhesion molecules (XMAD), the polynucleotides encoding XMAD, and the use of these compositions for the diagnosis, treatment, or prevention of genetic, autoimmune/inflammation, and cell proliferative disorders, including cancer.

[0129] Table 1 lists the Incyte clones used to assemble full length nucleotide sequences encoding XMAD. Columns 1 and 2 show the sequence identification numbers (SEQ ID NOs) of the polypeptide and nucleotide sequences, respectively. Column 3 shows the clone IDs of the Incyte clones in which nucleic acids encoding each XMAD were identified, and column 4 shows the cDNA libraries from which these clones were isolated. Column 5 shows Incyte clones and their corresponding cDNA libraries. Clones for which cDNA libraries are not indicated were derived from pooled cDNA libraries. In some cases, GenBank sequence identifiers are also shown in column 5. The Incyte clones and GenBank cDNA sequences, where indicated, in column 5 were used to assemble the consensus nucleotide sequence of each XMAD and are useful as fragments in hybridization technologies.

[0130] The columns of Table 2 show various properties of each of the polypeptides of the invention: column I references the SEQ ID NO; column 2 shows the number of amino acid residues in each polypeptide; column 3 shows potential phosphorylation sites; column 4 shows potential glycosylation sites; column 5 shows the amino acid residues comprising signature sequences and motifs; column 6 shows homologous sequences as identified by BLAST analysis along with relevant citations, all of which are expressly incorporated by reference herein in their entirety; and column 7 shows analytical methods and in some cases, searchable databases to which the analytical methods were applied. The methods of column 7 were used to characterize each polypeptide through sequence homology and protein motifs.

[0131] The columns of Table 3 show the tissue-specificity and diseases, disorders, or conditions associated with nucleotide sequences encoding XMAD. The first column of Table 3 lists the nucleotide SEQ ID NOs. Column 2 lists fragments of the nucleotide sequences of column 1. These fragments are useful, for example, in hybridization or amplification technologies to identify SEQ ID NO:22-42 and to distinguish between SEQ ID NO:22-42 and related polynucleotide sequences. The polypeptides encoded by these fragments are useful, for example, as immunogenic peptides. Column 3 lists tissue categories which express XMAD as a fraction of total tissues expressing XMAD. Column 4 lists diseases, disorders, or conditions associated with those tissues expressing XMAD as a fraction of total tissues expressing XMAD. Column 5 lists the vectors used to subclone each cDNA library.

[0132] The columns of Table 4 show descriptions of the tissues used to construct the cDNA libraries from which cDNA clones encoding XMAD were isolated. Column I references the nucleotide SEQ ID NOs, column 2 shows the cDNA libraries from which these clones were isolated, and column 3 shows the tissue origins and other descriptive information relevant to the cDNA libraries in column 2.

[0133] SEQ ID NO:18 maps to chromosome 22 within the interval from the P terminus to 19.5 centiMorgans.

[0134] The invention also encompasses XMAD variants. A preferred XMAD variant is one which has at least about 80%, or alternatively at least about 90%, or even at least about 95% amino acid sequence identity to the XMAD amino acid sequence, and which contains at least one functional or structural characteristic of XMAD.

[0135] The invention also encompasses polynucleotides which encode XMAD. In a particular embodiment, the invention encompasses a polynucleotide sequence comprising a sequence selected from the group consisting of SEQ ID NO:22-42, which encodes XMAD. The polynucleotide sequences of SEQ ID NO:22-42, as presented in the Sequence Listing, embrace the equivalent RNA sequences, wherein occurrences of the nitrogenous base thymine are replaced with uracil, and the sugar backbone is composed of ribose instead of deoxyribose.

[0136] The invention also encompasses a variant of a polynucleotide sequence encoding XMAD. In particular, such a variant polynucleotide sequence will have at least about 70%, or alternatively at least about 85%, or even at least about 95% polynucleotide sequence identity to the polynucleotide sequence encoding XMAD. A particular aspect of the invention encompasses a variant of a polynucleotide sequence comprising a sequence selected from the group consisting of SEQ ID NO:22-42 which has at least about 70%, or alternatively at least about 85%, or even at least about 95% polynucleotide sequence identity to a nucleic acid sequence selected from the group consisting of SEQ ID NO:22-42. Any one of the polynucleotide variants described above can encode an amino acid sequence which contains at least one functional or structural characteristic of XMAD.

[0137] It will be appreciated by those skilled in the art that as a result of the degeneracy of the genetic code, a multitude of polynucleotide sequences encoding XMAD, some bearing minimal similarity to the polynucleotide sequences of any known and naturally occurring gene, may be produced. Thus, the invention contemplates each and every possible variation of polynucleotide sequence that could be made by selecting combinations based on possible codon choices. These combinations are made in accordance with the standard triplet genetic code as applied to the polynucleotide sequence of naturally occurring XMAD, and all such variations are to be considered as being specifically disclosed.

[0138] Although nucleotide sequences which encode XMAD and its variants are generally capable of hybridizing to the nucleotide sequence of the naturally occurring XMAD under appropriately selected conditions of stringency, it may be advantageous to produce nucleotide sequences encoding XMAD or its derivatives possessing a substantially different codon usage, e.g., inclusion of non-naturally occurring codons. Codons may be selected to increase the rate at which expression of the peptide occurs in a particular prokaryotic or eukaryotic host in accordance with the frequency with which particular codons are utilized by the host. Other reasons for substantially altering the nucleotide sequence encoding XMAD and its derivatives without altering the encoded amino acid sequences include the production of RNA transcripts having more desirable properties, such as a greater half-life, than transcripts produced from the naturally occurring sequence.

[0139] The invention also encompasses production of DNA sequences which encode XMAD and XMAD derivatives, or fragments thereof, entirely by synthetic chemistry. After production, the synthetic sequence may be inserted into any of the many available expression vectors and cell systems using reagents well known in the art. Moreover, synthetic chemistry may be used to introduce mutations into a sequence encoding XMAD or any fragment thereof.

[0140] Also encompassed by the invention are polynucleotide sequences that are capable of hybridizing to the claimed polynucleotide sequences, and, in particular, to those shown in SEQ ID NO:22-42 and fragments thereof under various conditions of stringency. (See, e.g., Wahl, G. M. and S. L. Berger (1987) Methods Enzymol. 152:399-407; Kimmel, A. R. (1987) Methods Enzymol. 152:507-511.) Hybridization conditions, including annealing and wash conditions, are described in “Definitions.”

[0141] Methods for DNA sequencing are well known in the art and may be used to practice any of the embodiments of the invention. The methods may employ such enzymes as the Klenow fragment of DNA polymerase 1, SEQUENASE (US Biochemical, Cleveland Ohio), Taq polymerase (Applied Biosystems, Foster City Calif.), thermostable T7 polymerase (Amersham Pharmacia Biotech, Piscataway N.J.), or combinations of polymerases and proofreading exonucleases such as those found in the ELONGASE amplification system (Life Technologies, Gaithersburg Md.). Preferably, sequence preparation is automated with machines such as the MICROLAB 2200 liquid transfer system (Hamilton, Reno Nev.), PTC200 thermal cycler (MJ Research, Watertown Mass.) and ABI CATALYST 800 thermal cycler (Applied Biosystems). Sequencing is then carried out using either the ABI 373 or 377 DNA sequencing system (Applied Biosystems), the MEGABACE 1000 DNA sequencing system (Molecular Dynamics, Sunnyvale Calif.), or other systems known in the art. The resulting sequences are analyzed using a variety of algorithms which are well known in the art. (See, e.g., Ausubel, F. M. (1997) Short Protocols in Molecular Biology, John Wiley & Sons, New York N.Y., unit 7.7; Meyers, R. A. (1995) Molecular Biology and Biotechnology, Wiley VCH, New York N.Y., pp. 856-853.)

[0142] The nucleic acid sequences encoding XMAD may be extended utilizing a partial nucleotide sequence and employing various PCR-based methods known in the art to detect upstream sequences, such as promoters and regulatory elements. For example, one method which may be employed, restriction-site PCR, uses universal and nested primers to amplify unknown sequence from genomic DNA within a cloning vector. (See, e.g., Sarkar, G. (1993) PCR Methods Applic. 2:318-322.) Another method, inverse PCR, uses primers that extend in divergent directions to amplify unknown sequence from a circularized template. The template is derived from restriction fragments comprising a known genomic locus and surrounding sequences. (See, e.g., Triglia, T. et al. (1988) Nucleic Acids Res. 16:8186.) A third method, capture PCR, involves PCR amplification of DNA fragments adjacent to known sequences in human and yeast artificial chromosome DNA. (See, e.g., Lagerstrom, M. et al. (1991) PCR Methods Applic. 1:111-119.) In this method, multiple restriction enzyme digestions and ligations may be used to insert an engineered double-stranded sequence into a region of unknown sequence before performing PCR. Other methods which may be used to retrieve unknown sequences are known in the art. (See, e.g., Parker, J. D. et al. (1991) Nucleic Acids Res. 19:3055-3060). Additionally, one may use PCR, nested primers, and PROMOTERFINDER libraries (Clontech, Palo Alto Calif.) to walk genomic DNA. This procedure avoids the need to screen libraries and is useful in finding intron/exon junctions. For all PCR-based methods, primers may be designed using commercially available software, such as OLIGO 4.06 Primer Analysis software (National Biosciences, Plymouth Minn.) or another appropriate program, to be about 22 to 30 nucleotides in length, to have a GC content of about 50% or more, and to anneal to the template at temperatures of about 68° C. to 72° C.

[0143] When screening for full-length cDNAs, it is preferable to use libraries that have been size-selected to include larger cDNAs. In addition, random-primed libraries, which often include sequences containing the 5′ regions of genes, are preferable for situations in which an oligo d(T) library does not yield a full-length cDNA. Genomic libraries may be useful for extension of sequence into 5′ non-transcribed regulatory regions.

[0144] Capillary electrophoresis systems which are commercially available may be used to analyze the size or confirm the nucleotide sequence of sequencing or PCR products. In particular, capillary sequencing may employ flowable polymers for electrophoretic separation, four different nucleotide-specific, laser-stimulated fluorescent dyes, and a charge coupled device camera for detection of the emitted wavelengths. Output/light intensity may be converted to electrical signal using appropriate software (e.g., GENOTYPER and SEQUENCE NAVIGATOR, Applied Biosystems), and the entire process from loading of samples to computer analysis and electronic data display may be computer controlled. Capillary electrophoresis is especially preferable for sequencing small DNA fragments which may be present in limited amounts in a particular sample.

[0145] In another embodiment of the invention, polynucleotide sequences or fragments thereof which encode XMAD may be cloned in recombinant DNA molecules that direct expression of XMAD, or fragments or functional equivalents thereof, in appropriate host cells. Due to the inherent degeneracy of the genetic code, other DNA sequences which encode substantially the same or a functionally equivalent amino acid sequence may be produced and used to express XMAD.

[0146] The nucleotide sequences of the present invention can be engineered using methods generally known in the art in order to alter XMAD-encoding sequences for a variety of purposes including, but not limited to, modification of the cloning, processing, and/or expression of the gene product. DNA shuffling by random fragmentation and PCR reassembly of gene fragments and synthetic oligonucleotides may be used to engineer the nucleotide sequences. For example, oligonucleotide-mediated site-directed mutagenesis may be used to introduce mutations that create new restriction sites, alter glycosylation patterns, change codon preference, produce splice variants, and so forth.

[0147] The nucleotides of the present invention may be subjected to DNA shuffling techniques such as MOLECULARBREEDING (Maxygen Inc., Santa Clara Calif.; described in U.S. Pat. No. 5,837,458; Chang, C.-C. et al. (1999) Nat. Biotechnol. 17:793-797; Christians, F. C. et al. (1999) Nat. Biotechnol. 17:259-264; and Crameri, A. et al. (1996) Nat. Biotechnol. 14:315-319) to alter or improve the biological properties of XMAD, such as its biological or enzymatic activity or its ability to bind to other molecules or compounds. DNA shuffling is a process by which a library of gene variants is produced using PCR-mediated recombination of gene fragments. The library is then subjected to selection or screening procedures that identify those gene variants with the desired properties. These preferred variants may then be pooled and further subjected to recursive rounds of DNA shuffling and selection/screening. Thus, genetic diversity is created through “artificial” breeding and rapid molecular evolution. For example, fragments of a single gene containing random point mutations may be recombined, screened, and then reshuffled until the desired properties are optimized. Alternatively, fragments of a given gene may be recombined with fragments of homologous genes in the same gene family, either from the same or different species, thereby maximizing the genetic diversity of multiple naturally occurring genes in a directed and controllable manner.

[0148] In another embodiment, sequences encoding XMAD may be synthesized, in whole or in part, using chemical methods well known in the art. (See, e.g., Caruthers, M. H. et al. (1980) Nucleic Acids Symp. Ser. 7:215-223; Horn, T. et al. (1980) Nucleic Acids Symp. Ser. 7:225-232.) Alternatively, XMAD itself or a fragment thereof may be synthesized using chemical methods. For example, peptide synthesis can be performed using various solution-phase or solid-phase techniques. (See, e.g., Creighton, T. (1984) Proteins, Structures and Molecular Properties, W H Freeman, New York N.Y., pp. 55-60; and Roberge, J. Y. et al. (1995) Science 269:202-204.) Automated synthesis may be achieved using the ABI 431A peptide synthesizer (Applied Biosystems). Additionally, the amino acid sequence of XMAD, or any part thereof, may be altered during direct synthesis and/or combined with sequences from other proteins, or any part thereof, to produce a variant polypeptide or a polypeptide having a sequence of a naturally occurring polypeptide.

[0149] The peptide may be substantially purified by preparative high performance liquid chromatography. (See, e.g., Chiez, R. M. and F. Z. Regnier (1990) Methods Enzymol. 182:392-421.) The composition of the synthetic peptides may be confirmed by amino acid analysis or by sequencing. (See, e.g., Creighton, supra, pp. 28-53.)

[0150] In order to express a biologically active XMAD, the nucleotide sequences encoding XMAD or derivatives thereof may be inserted into an appropriate expression vector, i.e., a vector which contains the necessary elements for transcriptional and translational control of the inserted coding sequence in a suitable host. These elements include regulatory sequences, such as enhancers, constitutive and inducible promoters, and 5′ and 3′ untranslated regions in the vector and in polynucleotide sequences encoding XMAD. Such elements may vary in their strength and specificity. Specific initiation signals may also be used to achieve more efficient translation of sequences encoding XMAD. Such signals include the ATG initiation codon and adjacent sequences, e.g. the Kozak sequence. In cases where sequences encoding XMAD and its initiation codon and upstream regulatory sequences are inserted into the appropriate expression vector, no additional transcriptional or translational control signals may be needed. However, in cases where only coding sequence, or a fragment thereof, is inserted, exogenous translational control signals including an in-frame ATG initiation codon should be provided by the vector. Exogenous translational elements and initiation codons may be of various origins, both natural and synthetic. The efficiency of expression may be enhanced by the inclusion of enhancers appropriate for the particular host cell system used. (See, e.g., Scharf, D. et al. (1994) Results Probl. Cell Differ. 20:125-162.)

[0151] Methods which are well known to those skilled in the art may be used to construct expression vectors containing sequences encoding XMAD and appropriate transcriptional and translational control elements. These methods include in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. (See, e.g., Sambrook, J. et al. (1989) Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Press, Plainview N.Y., ch. 4, 8, and 16-17; Ausubel, F. M. et al. (1995) Current Protocols in Molecular Biology, John Wiley & Sons, New York N.Y., ch. 9, 13, and 16.)

[0152] A variety of expression vector/host systems may be utilized to contain and express sequences encoding XMAD. These include, but are not limited to, microorganisms such as bacteria transformed with recombinant bacteriophage, plasmid, or cosmid DNA expression vectors; yeast transformed with yeast expression vectors; insect cell systems infected with viral expression vectors (e.g., baculovirus); plant cell systems transformed with viral expression vectors (e.g., cauliflower mosaic virus, CaMV, or tobacco mosaic virus, TMV) or with bacterial expression vectors (e.g., Ti or pBR322 plasmids); or animal cell systems. (See, e.g., Sambrook, supra; Ausubel, supra; Van Heeke, G. and S. M. Schuster (1989) J. Biol. Chem. 264:5503-5509; Bitter, G. A. et al. (1987) Methods Enzymol. 153:516-544; Scorer, C. A. et al. (1994) Bio/Technology 12:181-184; Engelhard, E. K. et al. (1994) Proc. Natl. Acad. Sci. USA 91:3224-3227; Sandig, V. et al. (1996) Hum. Gene Ther. 7:1937-1945; Takamatsu, N. (1987) EMBO J. 6:307-311; Coruzzi, G. et al. (1984) EMBO J. 3:1671-1680; Broglie, R. et al. (1984) Science 224:838-843; Winter, J. et al. (1991) Results Probl. Cell Differ. 17:85-105; The McGraw Hill Yearbook of Science and Technology (1992) McGraw Hill, New York N.Y., pp. 191-196; Logan, J. and T. Shenk (1984) Proc. Natl. Acad. Sci. USA 81:3655-3659; and Harrington, J. J. et al., (1997) Nat. Genet. 15:345-355.) Expression vectors derived from retroviruses, adenoviruses, or herpes or vaccinia viruses, or from various bacterial plasmids, may be used for delivery of nucleotide sequences to the targeted organ, tissue, or cell population. (See, e.g., Di Nicola, M. et al., (1998) Cancer Gen. Ther. 5(6):350-356; Yu, M. et al. (1993) Proc. Natl. Acad. Sci. USA 90(13):6340-6344; Buller, R. M. et al. (1985) Nature 317(6040):813-815; McGregor, D. P. et al. (1994) Mol. Immunol. 31(3):219-226; and Verma, I. M. and N. Somia (1997) Nature 389:239-242.) The invention is not limited by the host cell employed.

[0153] In bacterial systems, a number of cloning and expression vectors may be selected depending upon the use intended for polynucleotide sequences encoding XMAD. For example, routine cloning, subcloning, and propagation of polynucleotide sequences encoding XMAD can be achieved using a multifunctional E. coli vector such as PBLUESCRIPT (Stratagene, La Jolla Calif.) or PSPORT1 plasmid (Life Technologies). Ligation of sequences encoding XMAD into the vector's multiple cloning site disrupts the lacZ gene, allowing a calorimetric screening procedure for identification of transformed bacteria containing recombinant molecules. In addition, these vectors may be useful for in vitro transcription, dideoxy sequencing, single strand rescue with helper phage, and creation of nested deletions in the cloned sequence. (See, e.g., Van Heeke, G. and S.M. Schuster (1989) J. Biol. Chem. 264:5503-5509.) When large quantities of XMAD are needed, e.g. for the production of antibodies, vectors which direct high level expression of XMAD may be used. For example, vectors containing the strong, inducible T5 or T7 bacteriophage promoter may be used.

[0154] Yeast expression systems may be used for production of XMAD. A number of vectors containing constitutive or inducible promoters, such as alpha factor, alcohol oxidase, and PGH promoters, may be used in the yeast Saccharomyces cerevisiae or Pichia pastoris. In addition, such vectors direct either the secretion or intracellular retention of expressed proteins and enable integration of foreign sequences into the host genome for stable propagation. (See, e.g., Ausubel, 1995, supra; Bitter, supra; and Scorer, supra.) Plant systems may also be used for expression of XMAD. Transcription of sequences encoding XMAD may be driven viral promoters, e.g., the 35S and 19S promoters of CaMV used alone or in combination with the omega leader sequence from TMV (Takamatsu, N. (1987) EMBO J. 6:307-311). Alternatively, plant promoters such as the small subunit of RUBISCO or heat shock promoters may be used. (See, e.g., Coruzzi, supra; Broglie, supra; and Winter, supra.) These constructs can be introduced into plant cells by direct DNA transformation or pathogen-mediated transfection. (See, e.g., The McGraw Hill Yearbook of Science and Technology (1992) McGraw Hill, New York N.Y., pp. 191-196.)

[0155] In mammalian cells, a number of viral-based expression systems may be utilized. In cases where an adenovirus is used as an expression vector, sequences encoding XMAD may be ligated into an adenovirus transcription/translation complex consisting of the late promoter and tripartite leader sequence. Insertion in a non-essential E1 or E3 region of the viral genome may be used to obtain infective virus which expresses XMAD in host cells. (See, e.g., Logan, J. and T. Shenk (1984) Proc. Natl. Acad. Sci. USA 81:3655-3659.) In addition, transcription enhancers, such as the Rous sarcoma virus (RSV) enhancer, may be used to increase expression in mammalian host cells. SV40 or EBV-based vectors may also be used for high-level protein expression.

[0156] Human artificial chromosomes (HACs) may also be employed to deliver larger fragments of DNA than can be contained in and expressed from a plasmid. HACs of about 6 kb to 10 Mb are constructed and delivered via conventional delivery methods (liposomes, polycationic amino polymers, or vesicles) for therapeutic purposes. (See, e.g., Harrington, J. J. et al. (1997) Nat. Genet. 15:345-355.)

[0157] For long term production of recombinant proteins in mammalian systems, stable expression of XMAD in cell lines is preferred. For example, sequences encoding XMAD can be transformed into cell lines using expression vectors which may contain viral origins of replication and/or endogenous expression elements and a selectable marker gene on the same or on a separate vector. Following the introduction of the vector, cells may be allowed to grow for about 1 to 2 days in enriched media before being switched to selective media. The purpose of the selectable marker is to confer resistance to a selective agent, and its presence allows growth and recovery of cells which successfully express the introduced sequences. Resistant clones of stably transformed cells may be propagated using tissue culture techniques appropriate to the cell type.

[0158] Any number of selection systems may be used to recover transformed cell lines. These include, but are not limited to, the herpes simplex virus thymidine kinase and adenine phosphoribosyltransferase genes, for use in tk⁻ and apr⁻ cells, respectively. (See, e.g., Wigler, M. et al. (1977) Cell 11:223-232; Lowy, I. et al. (1980) Cell 22:817-823.) Also, antimetabolite, antibiotic, or herbicide resistance can be used as the basis for selection. For example, dhfr confers resistance to methotrexate; neo confers resistance to the aminoglycosides neomycin and G418; and als and pat confer resistance to chlorsulfuron and phosphinotricin acetyltransferase, respectively. (See, e.g., Wigler, M. et al. (1980) Proc. Natl. Acad. Sci. USA 77:3567-3570; Colbere-Garapin, F. et al. (1981) J. Mol. Biol. 150:1-14.) Additional selectable genes have been described, e.g., trpb and hisD, which alter cellular requirements for metabolites. (See, e.g., Hartman, S. C. and R. C. Mulligan (1988) Proc. Natl. Acad. Sci. USA 85:8047-8051.) Visible markers, e.g., anthocyanins, green fluorescent proteins (GFP; Clontech), β glucuronidase and its substrate β-glucuronide, or luciferase and its substrate luciferin may be used. These markers can be used not only to identify transformants, but also to quantify the amount of transient or stable protein expression attributable to a specific vector system. (See, e.g., Rhodes, C. A. (1995) Methods Mol. Biol. 55:121-131.)

[0159] Although the presence/absence of marker gene expression suggests that the gene of interest is also present, the presence and expression of the gene may need to be confirmed. For example, if the sequence encoding XMAD is inserted within a marker gene sequence, transformed cells containing sequences encoding XMAD can be identified by the absence of marker gene function. Alternatively, a marker gene can be placed in tandem with a sequence encoding XMAD under the control of a single promoter. Expression of the marker gene in response to induction or selection usually indicates expression of the tandem gene as well.

[0160] In general, host cells that contain the nucleic acid sequence encoding XMAD and that express XMAD may be identified by a variety of procedures known to those of skill in the art. These procedures include, but are not limited to, DNA-DNA or DNA-RNA hybridizations, PCR amplification, and protein bioassay or immunoassay techniques which include membrane, solution, or chip based technologies for the detection and/or quantification of nucleic acid or protein sequences.

[0161] Immunological methods for detecting and measuring the expression of XMAD using either specific polyclonal or monoclonal antibodies are known in the art. Examples of such techniques include enzyme-linked immunosorbent assays (ELISAs), radioimmunoassays (RIAs), and fluorescence activated cell sorting (FACS). A two-site, monoclonal-based immunoassay utilizing monoclonal antibodies reactive to two non-interfering epitopes on XMAD is preferred, but a competitive binding assay may be employed. These and other assays are well known in the art. (See, e.g., Hampton, R. et al. (1990) Serological Methods, a Laboratory Manual, APS Press, St. Paul Minn., Sect. IV; Coligan, J. E. et al. (1997) Current Protocols in Immunology, Greene Pub. Associates and Wiley-Interscience, New York N.Y.; and Pound, J. D. (1998) Immunochemical Protocols, Humana Press, Totowa N.J.)

[0162] A wide variety of labels and conjugation techniques are known by those skilled in the art and may be used in various nucleic acid and amino acid assays. Means for producing labeled hybridization or PCR probes for detecting sequences related to polynucleotides encoding XMAD include oligolabeling, nick translation, end-labeling, or PCR amplification using a labeled nucleotide. Alternatively, the sequences encoding XMAD, or any fragments thereof, may be cloned into a vector for the production of an mRNA probe. Such vectors are known in the art, are commercially available, and may be used to synthesize RNA probes in vitro by addition of an appropriate RNA polymerase such as T7, T3, or SP6 and labeled nucleotides. These procedures may be conducted using a variety of commercially available kits, such as those provided by Amersham Pharmacia Biotech, Promega (Madison Wis.), and US Biochemical. Suitable reporter molecules or labels which may be used for ease of detection include radionuclides, enzymes, fluorescent, chemiluminescent, or chromogenic agents, as well as substrates, cofactors, inhibitors, magnetic particles, and the like.

[0163] Host cells transformed with nucleotide sequences encoding XMAD may be cultured under conditions suitable for the expression and recovery of the protein from cell culture. The protein produced by a transformed cell may be secreted or retained intracellularly depending on the sequence and/or the vector used. As will be understood by those of skill in the art, expression vectors containing polynucleotides which encode XMAD may be designed to contain signal sequences which direct secretion of XMAD through a prokaryotic or eukaryotic cell membrane.

[0164] In addition, a host cell strain may be chosen for its ability to modulate expression of the inserted sequences or to process the expressed protein in the desired fashion. Such modifications of the polypeptide include, but are not limited to, acetylation, carboxylation, glycosylation, phosphorylation, lipidation, and acylation. Post-translational processing which cleaves a “prepro” or “pro” form of the protein may also be used to specify protein targeting, folding, and/or activity. Different host cells which have specific cellular machinery and characteristic mechanisms for post-translational activities (e.g., CHO, HeLa, MDCK, HEK293, and WI38) are available from the American Type Culture Collection (ATCC, Manassas Va.) and may be chosen to ensure the correct modification and processing of the foreign protein.

[0165] In another embodiment of the invention, natural, modified, or recombinant nucleic acid sequences encoding XMAD may be ligated to a heterologous sequence resulting in translation of a fusion protein in any of the aforementioned host systems. For example, a chimeric XMAD protein containing a heterologous moiety that can be recognized by a commercially available antibody may facilitate the screening of peptide libraries for inhibitors of XMAD activity. Heterologous protein and peptide moieties may also facilitate purification of fusion proteins using commercially available affinity matrices. Such moieties include, but are not limited to, glutathione S-transferase (GST), maltose binding protein (MBP), thioredoxin (Trx), calmodulin binding peptide (CBP), 6-His, FLAG, c-myc, and hemagglutinin (HA). GST, MBP, Trx, CBP, and 6-His enable purification of their cognate fusion proteins on immobilized glutathione, maltose, phenylarsine oxide, calmodulin, and metal-chelate resins, respectively. FLAG, c-myc, and hemagglutinin (HA) enable immunoaffinity purification of fusion proteins using commercially available monoclonal and polyclonal antibodies that specifically recognize these epitope tags. A fusion protein may also be engineered to contain a proteolytic cleavage site located between the XMAD encoding sequence and the heterologous protein sequence, so that XMAD may be cleaved away from the heterologous moiety following purification. Methods for fusion protein expression and purification are discussed in Ausubel (1995, supra, ch. 10). A variety of commercially available kits may also be used to facilitate expression and purification of fusion proteins.

[0166] In a further embodiment of the invention, synthesis of radiolabeled XMAD may be achieved in vitro using the TNT rabbit reticulocyte lysate or wheat germ extract system (Promega). These systems couple transcription and translation of protein-coding sequences operably associated with the T7, T3, or SP6 promoters. Translation takes place in the presence of a radiolabeled amino acid precursor, for example, ³⁵S-methionine.

[0167] XMAD of the present invention or fragments thereof may be used to screen for compounds that specifically bind to XMAD. At least one and up to a plurality of test compounds may be screened for specific binding to XMAD. Examples of test compounds include antibodies, oligonucleotides, proteins (e.g., receptors), or small molecules.

[0168] In one embodiment, the compound thus identified is closely related to the natural ligand of XMAD, e.g., a ligand or fragment thereof, a natural substrate, a structural or functional mimetic, or a natural binding partner. (See, e.g., Coligan, J. E. et al. (1991) Current Protocols in Immunology 1(2): Chapter 5.) Similarly, the compound can be closely related to the natural receptor to which XMAD binds, or to at least a fragment of the receptor, e.g., the ligand binding site. In either case, the compound can be rationally designed using known techniques. In one embodiment, screening for these compounds involves producing appropriate cells which express XMAD, either as a secreted protein or on the cell membrane. Preferred cells include cells from mammals, yeast, Drosophila, or E. coli. Cells expressing XMAD or cell membrane fractions which contain XMAD are then contacted with a test compound and binding, stimulation, or inhibition of activity of either XMAD or the compound is analyzed.

[0169] An assay may simply test binding of a test compound to the polypeptide, wherein binding is detected by a fluorophore, radioisotope, enzyme conjugate, or other detectable label. For example, the assay may comprise the steps of combining at least one test compound with XMAD, either in solution or affixed to a solid support, and detecting the binding of XMAD to the compound. Alternatively, the assay may detect or measure binding of a test compound in the presence of a labeled competitor. Additionally, the assay may be carried out using cell-free preparations, chemical libraries, or natural product mixtures, and the test compound(s) may be free in solution or affixed to a solid support.

[0170] XMAD of the present invention or fragments thereof may be used to screen for compounds that modulate the activity of XMAD. Such compounds may include agonists, antagonists, or partial or inverse agonists. In one embodiment, an assay is performed under conditions permissive for XMAD activity, wherein XMAD is combined with at least one test compound, and the activity of XMAD in the presence of a test compound is compared with the activity of XMAD in the absence of the test compound. A change in the activity of XMAD in the presence of the test compound is indicative of a compound that modulates the activity of XMAD. Alternatively, a test compound is combined with an in vitro or cell-free system comprising XMAD under conditions suitable for XMAD activity, and the assay is performed. In either of these assays, a test compound which modulates the activity of XMAD may do so indirectly and need not come in direct contact with the test compound. At least one and up to a plurality of test compounds may be screened.

[0171] In another embodiment, polynucleotides encoding XMAD or their mammalian homologs may be “knocked out” in an animal model system using homologous recombination in embryonic stem (ES) cells. Such techniques are well known in the art and are useful for the generation of animal models of human disease. (See, e.g., U.S. Pat. No. 5,175,383 and U.S. Pat. No. 5,767,337.) For example, mouse ES cells, such as the mouse 129/SvJ cell line, are derived from the early mouse embryo and grown in culture. The ES cells are transformed with a vector containing the gene of interest disrupted by a marker gene, e.g., the neomycin phosphotransferase gene (neo; Capecchi, M. R. (1989) Science 244:1288-1292). The vector integrates into the corresponding region of the host genome by homologous recombination. Alternatively, homologous recombination takes place using the Cre-loxP system to knockout a gene of interest in a tissue- or developmental stage-specific manner (Marth, J. D. (1996) Clin. Invest. 97:1999-2002; Wagner, K. U. et al. (1997) Nucleic Acids Res. 25:4323-4330). Transformed ES cells are identified and microinjected into mouse cell blastocysts such as those from the C57BL/6 mouse strain. The blastocysts are surgically transferred to pseudopregnant dams, and the resulting chimeric progeny are genotyped and bred to produce heterozygous or homozygous strains. Transgenic animals thus generated may be tested with potential therapeutic or toxic agents.

[0172] Polynucleotides encoding XMAD may also be manipulated in vitro in ES cells derived from human blastocysts. Human ES cells have the potential to differentiate into at least eight separate cell lineages including endoderm, mesoderm, and ectodermal cell types. These cell lineages differentiate into, for example, neural cells, hematopoietic lineages, and cardiomyocytes (Thomson, J. A. et al. (1998) Science 282:1145-1147).

[0173] Polynucleotides encoding XMAD can also be used to create “knockin” humanized animals (pigs) or transgenic animals (mice or rats) to model human disease. With knockin technology, a region of a polynucleotide encoding XMAD is injected into animal ES cells, and the injected sequence integrates into the animal cell genome. Transformed cells are injected into blastulae, and the blastulae are implanted as described above. Transgenic progeny or inbred lines are studied and treated with potential pharmaceutical agents to obtain information on treatment of a human disease. Alternatively, a mammal inbred to overexpress XMAD, e.g., by secreting XMAD in its milk, may also serve as a convenient source of that protein (Janne, J. et al. (1998) Biotechnol. Annu. Rev. 4:55-74).

[0174] Therapeutics

[0175] Chemical and structural similarity, e.g., in the context of sequences and motifs, exists between regions of XMAD and extracellular matrix and cell adhesion molecules. In addition, the expression of XMAD is closely associated with cell proliferation. Therefore, XMAD appears to play a role in genetic, autoimmune/inflammation, and cell proliferative disorders, including cancer. In the treatment of disorders associated with increased XMAD expression or activity, it is desirable to decrease the expression or activity of XMAD. In the treatment of disorders associated with decreased XMAD expression or activity, it is desirable to increase the expression or activity of XMAD.

[0176] Therefore, in one embodiment, XMAD or a fragment or derivative thereof may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of XMAD. Examples of such disorders include, but are not limited to, a genetic disorder, such as adrenoleukodystrophy, Alport's syndrome, choroideremia, Duchenne and Becker muscular dystrophy, Down's syndrome, cystic fibrosis, chronic granulomatous disease, Gaucher's disease, Huntington's chorea, Marfan's syndrome, muscular dystrophy, myotonic dystrophy, pycnodysostosis, Refsum's syndrome, retinoblastoma, sickle cell anemia, thalassemia, Werner syndrome, von Willebrand's disease, Wilms' tumor, Zellweger syndrome, peroxisomal acyl-CoA oxidase deficiency, peroxisomal thiolase deficiency, peroxisomal bifunctional protein deficiency, mitochondrial carnitine pal mitoyl transferase and carnitine deficiency, mitochondrial very-long-chain acyl-CoA dehydrogenase deficiency, mitochondrial medium-chain acyl-CoA dehydrogenase deficiency, mitochondrial short-chain acyl-CoA dehydrogenase deficiency, mitochondrial electron transport flavoprotein and electron transport flavoprotein:ubiquinone oxidoreductase deficiency, mitochondrial trifunctional protein deficiency, and mitochondrial short-chain 3-hydroxyacyl-CoA dehydrogenase deficiency; an autoimmune/inflammation disorder, such as acquired immunodeficiency syndrome (AIDS), Addison's disease, adult respiratory distress syndrome, allergies, ankylosing spondylitis, amyloidosis, anemia, asthma, atherosclerosis, autoimmune hemolytic anemia, autoimmune thyroiditis, autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED), bronchitis, cholecystitis, contact dermatitis, Crohn's disease, atopic dermatitis, dermatomyositis, diabetes mellitus, emphysema, episodic lymphopenia with lymphocytotoxins, erythroblastosis fetalis, erythema nodosum, atrophic gastritis, glomerulonephritis, Goodpasture's syndrome, gout, Graves' disease, Hashimoto's thyroiditis, hypereosinophilia, irritable bowel syndrome, multiple sclerosis, myasthenia gravis, myocardial or pericardial inflammation, osteoarthritis, osteoporosis, pancreatitis, polymyositis, psoriasis, Reiter's syndrome, rheumatoid arthritis, scleroderma, Sjögren's syndrome, systemic anaphylaxis, systemic lupus erythematosus, systemic sclerosis, thrombocytopenic purpura, ulcerative colitis, uveitis, Werner syndrome, complications of cancer, hemodialysis, and extracorporeal circulation, viral, bacterial, fungal, parasitic, protozoal, and helminthic infections, and trauma; and a cell proliferative disorder, such as actinic keratosis, arteriosclerosis, atherosclerosis, bursitis, cirrhosis, hepatitis, mixed connective tissue disease (MCTD), myelofibrosis, paroxysmal nocturnal hemoglobinuria, polycythemia vera, psoriasis, primary thrombocythemia, and cancers including adenocarcinoma, leukemia, lymphoma, melanoma, myeloma, sarcoma, teratocarcinoma, and, in particular, cancers of the adrenal gland, bladder, bone, bone marrow, brain, breast, cervix, gall bladder, ganglia, gastrointestinal tract, heart, kidney, liver, lung, muscle, ovary, pancreas, parathyroid, penis, prostate, salivary glands, skin, spleen, testis, thymus, thyroid, and uterus.

[0177] In another embodiment, a vector capable of expressing XMAD or a fragment or derivative thereof may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of XMAD including, but not limited to, those described above.

[0178] In a further embodiment, a composition comprising a substantially purified XMAD in conjunction with a suitable pharmaceutical carrier may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of XMAD including, but not limited to, those provided above.

[0179] In still another embodiment, an agonist which modulates the activity of XMAD may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of XMAD including, but not limited to, those listed above.

[0180] In a further embodiment, an antagonist of XMAD may be administered to a subject to treat or prevent a disorder associated with increased expression or activity of XMAD. Examples of such disorders include, but are not limited to, those genetic, autoimmune/inflammation, and cell proliferative disorders, including cancer, described above. In one aspect, an antibody which specifically binds XMAD may be used directly as an antagonist or indirectly as a targeting or delivery mechanism for bringing a pharmaceutical agent to cells or tissues which express XMAD.

[0181] In an additional embodiment, a vector expressing the complement of the polynucleotide encoding XMAD may be administered to a subject to treat or prevent a disorder associated with increased expression or activity of XMAD including, but not limited to, those described above.

[0182] In other embodiments, any of the proteins, antagonists, antibodies, agonists, complementary sequences, or vectors of the invention may be administered in combination with other appropriate therapeutic agents. Selection of the appropriate agents for use in combination therapy may be made by one of ordinary skill in the art, according to conventional pharmaceutical principles. The combination of therapeutic agents may act synergistically to effect the treatment or prevention of the various disorders described above. Using this approach, one may be able to achieve therapeutic efficacy with lower dosages of each agent, thus reducing the potential for adverse side effects.

[0183] An antagonist of XMAD may be produced using methods which are generally known in the art. In particular, purified XMAD may be used to produce antibodies or to screen libraries of pharmaceutical agents to identify those which specifically bind XMAD. Antibodies to XMAD may also be generated using methods that are well known in the art. Such antibodies may include, but are not limited to, polyclonal, monoclonal, chimeric, and single chain antibodies, Fab fragments, and fragments produced by a Fab expression library. Neutralizing antibodies (i.e., those which inhibit dimer formation) are generally preferred for therapeutic use.

[0184] For the production of antibodies, various hosts including goats, rabbits, rats, mice, humans, and others may be immunized by injection with XMAD or with any fragment or oligopeptide thereof which has immunogenic properties. Depending on the host species, various adjuvants may be used to increase immunological response. Such adjuvants include, but are not limited to, Freund's, mineral gels such as aluminum hydroxide, and surface active substances such as lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, KLH, and dinitrophenol. Among adjuvants used in humans, BCG (bacilli Calmette-Guerin) and Corynebacterium parvum are especially preferable.

[0185] It is preferred that the oligopeptides, peptides, or fragments used to induce antibodies to XMAD have an amino acid sequence consisting of at least about 5 amino acids, and generally will consist of at least about 10 amino acids. It is also preferable that these oligopeptides, peptides, or fragments are identical to a portion of the amino acid sequence of the natural protein. Short stretches of XMAD amino acids may be fused with those of another protein, such as KLH, and antibodies to the chimeric molecule may be produced.

[0186] Monoclonal antibodies to XMAD may be prepared using any technique which provides for the production of antibody molecules by continuous cell lines in culture. These include, but are not limited to, the hybridoma technique, the human B-cell hybridoma technique, and the EBV-hybridoma technique. (See, e.g., Kohler, G. et al. (1975) Nature 256:495-497; Kozbor, D. et al. (1985) J. Immunol. Methods 81:3142; Cote, R. J. et al. (1983) Proc. Natl. Acad. Sci. USA 80:2026-2030; and Cole, S. P. et al. (1984) Mol. Cell Biol. 62:109-120.)

[0187] In addition, techniques developed for the production of “chimeric antibodies,” such as the splicing of mouse antibody genes to human antibody genes to obtain a molecule with appropriate antigen specificity and biological activity, can be used. (See, e.g., Morrison, S. L. et al. (1984) Proc. Natl. Acad. Sci. USA 81:6851-6855; Neuberger, M. S. et al. (1984) Nature 312:604-608; and Takeda, S. et al. (1985) Nature 314:452-454.) Alternatively, techniques described for the production of single chain antibodies may be adapted, using methods known in the art, to produce XMAD-specific single chain antibodies. Antibodies with related specificity, but of distinct idiotypic composition, may be generated by chain shuffling from random combinatorial immunoglobulin libraries. (See, e.g., Burton, D. R. (1991) Proc. Natl. Acad. Sci. USA 88:10134-10137.)

[0188] Antibodies may also be produced by inducing in vivo production in the lymphocyte population or by screening immunoglobulin libraries or panels of highly specific binding reagents as disclosed in the literature. (See, e.g., Orlandi, R. et al. (1989) Proc. Natl. Acad. Sci. USA 86:3833-3837; Winter, G. et al. (1991) Nature 349:293-299.)

[0189] Antibody fragments which contain specific binding sites for XMAD may also be generated. For example, such fragments include, but are not limited to, F(ab′)₂ fragments produced by pepsin digestion of the antibody molecule and Fab fragments generated by reducing the disulfide bridges of the F(ab′)₂ fragments. Alternatively, Fab expression libraries may be constructed to allow rapid and easy identification of monoclonal Fab fragments with the desired specificity. (See, e.g., Huse, W. D. et al. (1989) Science 246:1275-1281.)

[0190] Various immunoassays may be used for screening to identify antibodies having the desired specificity. Numerous protocols for competitive binding or immunoradiometric assays using either polyclonal or monoclonal antibodies with established specificities are well known in the art. Such immunoassays typically involve the measurement of complex formation between XMAD and its specific antibody. A two-site, monoclonal-based immunoassay utilizing monoclonal antibodies reactive to two non-interfering XMAD epitopes is generally used, but a competitive binding assay may also be employed (Pound, supra).

[0191] Various methods such as Scatchard analysis in conjunction with radioimmunoassay techniques may be used to assess the affinity of antibodies for XMAD. Affinity is expressed as an association constant, K_(a), which is defined as the molar concentration of XMAD-antibody complex divided by the molar concentrations of free antigen and free antibody under equilibrium conditions. The K_(a) determined for a preparation of polyclonal antibodies, which are heterogeneous in their affinities for multiple XMAD epitopes, represents the average affinity, or avidity, of the antibodies for XMAD. The K_(a) determined for a preparation of monoclonal antibodies, which are monospecific for a particular XMAD epitope, represents a true measure of affinity. High-affinity antibody preparations with K_(a) ranging from about 109 to 1012 L/mole are preferred for use in immunoassays in which the XMAD-antibody complex must withstand rigorous manipulations. Low-affinity antibody preparations with K_(a) ranging from about 10⁶ to 10⁷ L/mole are preferred for use in immunopurification and similar procedures which ultimately require dissociation of XMAD, preferably in active form, from the antibody (Catty, D. (1988) Antibodies, Volume I: A Practical Approach, IRL Press, Washington D.C.; Liddell, J. E. and A. Cryer (1991) A Practical Guide to Monoclonal Antibodies, John Wiley & Sons, New York N.Y.).

[0192] The titer and avidity of polyclonal antibody preparations may be further evaluated to determine the quality and suitability of such preparations for certain downstream applications. For example, a polyclonal antibody preparation containing at least 1-2 mg specific antibody/ml, preferably 5-10 mg specific antibody/ml, is generally employed in procedures requiring precipitation of XMAD-antibody complexes. Procedures for evaluating antibody specificity, titer, and avidity, and guidelines for antibody quality and usage in various applications, are generally available. (See, e.g., Catty, supra, and Coligan et al., supra.)

[0193] In another embodiment of the invention, the polynucleotides encoding XMAD, or any fragment or complement thereof, may be used for therapeutic purposes. In one aspect, modifications of gene expression can be achieved by designing complementary sequences or antisense molecules (DNA, RNA, PNA, or modified oligonucleotides) to the coding or regulatory regions of the gene encoding XMAD. Such technology is well known in the art, and antisense oligonucleotides or larger fragments can be designed from various locations along the coding or control regions of sequences encoding XMAD. (See, e.g., Agrawal, S., ed. (1996) Antisense Therapeutics, Humana Press Inc., Totawa N.J.)

[0194] In therapeutic use, any gene delivery system suitable for introduction of the antisense sequences into appropriate target cells can be used. Antisense sequences can be delivered intracellularly in the form of an expression plasmid which, upon transcription, produces a sequence complementary to at least a portion of the cellular sequence encoding the target protein. (See, e.g., Slater, J. E. et al. (1998) J. Allergy Clin. Immunol. 102(3):469-475; and Scanlon, K. J. et al. (1995) 9(13): 1288-1296.) Antisense sequences can also be introduced intracellularly through the use of viral vectors, such as retrovirus and adeno-associated virus vectors. (See, e.g., Miller, A. D. (1990) Blood 76:271; Ausubel, supra; Uckert, W. and W. Walther (1994) Pharmacol. Ther. 63(3):323-347.) Other gene delivery mechanisms include liposome-derived systems, artificial viral envelopes, and other systems known in the art. (See, e.g., Rossi, J. J. (1995) Br. Med. Bull. 51(1):217-225; Boado, R. J. et al (1998) J. Pharm. Sci. 87(11):1308-1315; and Morris, M. C. et al. (1997) Nucleic Acids Res. 25(14):2730-2736.)

[0195] In another embodiment of the invention, polynucleotides encoding XMAD may be used for somatic or germline gene therapy. Gene therapy may be performed to (i) correct a genetic deficiency (e.g., in the cases of severe combined immunodeficiency (SCID)-X1 disease characterized by X-linked inheritance (Cavazzana-Calvo, M. et al. (2000) Science 288:669-672), severe combined immunodeficiency syndrome associated with an inherited adenosine deaminase (ADA) deficiency (Blaese, R. M. et al. (1995) Science 270:475-480; Bordignon, C. et al. (1995) Science 270:470-475), cystic fibrosis (Zabner, J. et al. (1993) Cell 75:207-216; Crystal, R. G. et al. (1995) Hum. Gene Therapy 6:643-666; Crystal, R. G. et al. (1995) Hum. Gene Therapy 6:667-703), thalassamias, familial hypercholesterolemia, and hemophilia resulting from Factor VIII or Factor IX deficiencies (Crystal, R. G. (1995) Science 270:404-410; Verma, I. M. and N. Somia (1997) Nature 389:239-242)), (ii) express a conditionally lethal gene product (e.g., in the case of cancers which result from unregulated cell proliferation), or (iii) express a protein which affords protection against intracellular parasites (e.g., against human retroviruses, such as human immunodeficiency virus (HIV) (Baltimore, D. (1988) Nature 335:395-396; Poeschla, E. et al. (1996) Proc. Natl. Acad. Sci. USA. 93:11395-11399), hepatitis B or C virus (HBV, HCV); fungal parasites, such as Candida albicans and Paracoccidioides brasiliensis; and protozoan parasites such as Plasmodium falciparum and Trypanosoma cruzi). In the case where a genetic deficiency in XMAD expression or regulation causes disease, the expression of MAD from an appropriate population of transduced cells may alleviate the clinical manifestations caused by the genetic deficiency.

[0196] In a further embodiment of the invention, diseases or disorders caused by deficiencies in XMAD are treated by constructing mammalian expression vectors encoding XMAD and introducing these vectors by mechanical means into XMAD-deficient cells. Mechanical transfer technologies for use with cells in vivo or ex vitro include (i) direct DNA microinjection into individual cells, (ii) ballistic gold particle delivery, (iii) liposome-mediated transfection, (iv) receptor-mediated gene transfer, and (v) the use of DNA transposons (Morgan, R. A. and W. F. Anderson (1993) Annu. Rev. Biochem. 62:191-217; Ivics, Z. (1997) Cell 91:501-510; Boulay, J-L. and H. Récipon (1998) Curr. Opin. Biotechnol. 9:445-450).

[0197] Expression vectors that may be effective for the expression of XMAD include, but are not limited to, the PCDNA 3.1, EPITAG, PRCCMV2, PREP, PVAX vectors (Invitrogen, Carlsbad Calif.), PCMV-SCRIPT, PCMV-TAG, PEGSH/PERV (Stratagene, La Jolla Calif.), and PTET-OFF, PTET-ON, PTRE2, PTRE2-LUC, PTK-HYG (Clontech, Palo Alto Calif.). XMAD may be expressed using (i) a constitutively active promoter, (e.g., from cytomegalovirus (CMV), Rous sarcoma virus (RSV), SV40 virus, thymidine kinase (TK), or β-actin genes), (ii) an inducible promoter (e.g., the tetracycline-regulated promoter (Gossen, M. and H. Bujard (1992) Proc. Natl. Acad. Sci. USA 89:5547-5551; Gossen, M. et al. (1995) Science 268:1766-1769; Rossi, F. M. V. and H. M. Blau (1998) Curr. Opin. Biotechnol. 9:451-456), commercially available in the T-REX plasmid (Invitrogen)); the ecdysone-inducible promoter (available in the plasmids PVGRXR and PIND; Invitrogen); the FK506/rapamycin inducible promoter; or the RU486/mifepristone inducible promoter (Rossi, F. M. V. and H. M. Blau, supra)), or (iii) a tissue-specific promoter or the native promoter of the endogenous gene encoding XMAD from a normal individual.

[0198] Commercially available liposome transformation kits (e.g., the PERFECT LIPID TRANSFECTION KIT, available from Invitrogen) allow one with ordinary skill in the art to deliver polynucleotides to target cells in culture and require minimal effort to optimize experimental parameters. In the alternative, transformation is performed using the calcium phosphate method (Graham, F. L. and A. J. Eb (1973) Virology 52:456-467), or by electroporation (Neumann, E. et al. (1982) EMBO J. 1:841-845). The introduction of DNA to primary cells requires modification of these standardized mammalian transfection protocols.

[0199] In another embodiment of the invention, diseases or disorders caused by genetic defects with respect to XMAD expression are treated by constructing a retrovirus vector consisting of (i) the polynucleotide encoding XMAD under the control of an independent promoter or the retrovirus long terminal repeat (LTR) promoter, (ii) appropriate RNA packaging signals, and (iii) a Rev-responsive element (RRE) along with additional retrovirus cis-acting RNA sequences and coding sequences required for efficient vector propagation. Retrovirus vectors (e.g., PFB and PFBNEO) are commercially available (Stratagene) and are based on published data (Riviere, I. et al. (1995) Proc. Natl. Acad. Sci. USA 92:6733-6737), incorporated by reference herein. The vector is propagated in an appropriate vector producing cell line (VPCL) that expresses an envelope gene with a tropism for receptors on the target cells or a promiscuous envelope protein such as VSVg (Armentano, D. et al. (1987) J. Virol. 61:1647-1650; Bender, M. A. et al. (1987) J. Virol. 61:1639-1646; Adam, M. A. and A. D. Miller (1988) J. Virol. 62:3802-3806; Dull, T. et al. (1998) J. Virol. 72:8463-8471; Zufferey, R. et al. (1998) J. Virol. 72:9873-9880). U.S. Pat. No. 5,910,434 to Rigg (“Method for obtaining retrovirus packaging cell lines producing high transducing efficiency retroviral supernatant”) discloses a method for obtaining retrovirus packaging cell lines and is hereby incorporated by reference. Propagation of retrovirus vectors, transduction of a population of cells (e.g., CD4⁺ T-cells), and the return of transduced cells to a patient are procedures well known to persons skilled in the art of gene therapy and have been well documented (Ranga, U. et al. (1997) J. Virol. 71:7020-7029; Bauer, G. et al. (1997) Blood 89:2259-2267; Bonyhadi, M. L. (1997) J. Virol. 71:47074716; Ranga, U. et al. (1998) Proc. Natl. Acad. Sci. USA 95:1201-1206; Su, L. (1997) Blood 89:2283-2290).

[0200] In the alternative, an adenovirus-based gene therapy delivery system is used to deliver polynucleotides encoding XMAD to cells which have one or more genetic abnormalities with respect to the expression of XMAD. The construction and packaging of adenovirus-based vectors are well known to those with ordinary skill in the art. Replication defective adenovirus vectors have proven to be versatile for importing genes encoding immunoregulatory proteins into intact islets in the pancreas (Csete, M. E. et al. (1995) Transplantation 27:263-268). Potentially useful adenoviral vectors are described in U.S. Pat. No. 5,707,618 to Armentano (“Adenovirus vectors for gene therapy”), hereby incorporated by reference. For adenoviral vectors, see also Antinozzi, P. A. et al. (1999) Annu. Rev. Nutr. 19:511-544; and Verma, I. M. and N. Somia (1997) Nature 18:389:239-242, both incorporated by reference herein.

[0201] In another alternative, a herpes-based, gene therapy delivery system is used to deliver polynucleotides encoding XMAD to target cells which have one or more genetic abnormalities with respect to the expression of XMAD. The use of herpes simplex virus (HSV)-based vectors may be especially valuable for introducing XMAD to cells of the central nervous system, for which HSV has a tropism. The construction and packaging of herpes-based vectors are well known to those with ordinary skill in the art. A replication-competent herpes simplex virus (HSV) type 1-based vector has been used to deliver a reporter gene to the eyes of primates (Liu, X. et al. (1999) Exp. Eye Res. 169:385-395). The construction of a HSV-1 virus vector has also been disclosed in detail in U.S. Pat. No. 5,804,413 to DeLuca (“Herpes simplex virus strains for gene transfer”), which is hereby incorporated by reference. U.S. Pat. No. 5,804,413 teaches the use of recombinant HSV d92 which consists of a genome containing at least one exogenous gene to be transferred to a cell under the control of the appropriate promoter for purposes including human gene therapy. Also taught by this patent are the construction and use of recombinant HSV strains deleted for ICP4, ICP27 and ICP22. For HSV vectors, see also Goins, W. F. et al. (1999) J. Virol. 73:519-532 and Xu, H. et al. (1994) Dev. Biol. 163:152-161, hereby incorporated by reference. The manipulation of cloned herpesvirus sequences, the generation of recombinant virus following the transfection of multiple plasmids containing different segments of the large herpesvirus genomes, the growth and propagation of herpesvirus, and the infection of cells with herpesvirus are techniques well known to those of ordinary skill in the art.

[0202] In another alternative, an alphavirus (positive, single-stranded RNA virus) vector is used to deliver polynucleotides encoding XMAD to target cells. The biology of the prototypic alphavirus, Semliki Forest Virus (SFV), has been studied extensively and gene transfer vectors have been based on the SFV genome (Garoff, H. and K.-J. Li (1998) Curr. Opin. Biotechnol. 9:464-469). During alphavirus RNA replication, a subgenomic RNA is generated that normally encodes the viral capsid proteins. This subgenomic RNA replicates to higher levels than the full-length genomic RNA, resulting in the overproduction of capsid proteins relative to the viral proteins with enzymatic activity (e.g., protease and polymerase). Similarly, inserting the coding sequence for XMAD into the alphavirus genome in place of the capsid-coding region results in the production of a large number of XMAD-coding RNAs and the synthesis of high levels of XMAD in vector transduced cells. While alphavirus infection is typically associated with cell lysis within a few days, the ability to establish a persistent infection in hamster normal kidney cells (BHK-21) with a variant of Sindbis virus (SIN) indicates that the lytic replication of alphaviruses can be altered to suit the needs of the gene therapy application (Dryga, S. A. et al. (1997) Virology 228:74-83). The wide host range of alphaviruses will allow the introduction of XMAD into a variety of cell types. The specific transduction of a subset of cells in a population may require the sorting of cells prior to transduction. The methods of manipulating infectious cDNA clones of alphaviruses, performing alphavirus cDNA and RNA transfections, and performing alphavirus infections, are well known to those with ordinary skill in the art.

[0203] Oligonucleotides derived from the transcription initiation site, e.g., between about positions −10 and +10 from the start site, may also be employed to inhibit gene expression. Similarly, inhibition can be achieved using triple helix base-pairing methodology. Triple helix pairing is useful because it causes inhibition of the ability of the double helix to open sufficiently for the binding of polymerases, transcription factors, or regulatory molecules. Recent therapeutic advances using triplex DNA have been described in the literature. (See, e.g., Gee, J. E. et al. (1994) in Huber, B. E. and B. I. Carr, Molecular and Immunologic Approaches, Futura Publishing, Mt. Kisco N.Y., pp. 163-177.) A complementary sequence or antisense molecule may also be designed to block translation of mRNA by preventing the transcript from binding to ribosomes.

[0204] Ribozymes, enzymatic RNA molecules, may also be used to catalyze the specific cleavage of RNA. The mechanism of ribozyme action involves sequence-specific hybridization of the ribozyme molecule to complementary target RNA, followed by endonucleolytic cleavage. For example, engineered hammerhead motif ribozyme molecules may specifically and efficiently catalyze endonucleolytic cleavage of sequences encoding XMAD.

[0205] Specific ribozyme cleavage sites within any potential RNA target are initially identified by scanning the target molecule for ribozyme cleavage sites, including the following sequences: GUA, GUU, and GUC. Once identified, short RNA sequences of between 15 and 20 ribonucleotides, corresponding to the region of the target gene containing the cleavage site, may be evaluated for secondary structural features which may render the oligonucleotide inoperable. The suitability of candidate targets may also be evaluated by testing accessibility to hybridization with complementary oligonucleotides using ribonuclease protection assays.

[0206] Complementary ribonucleic acid molecules and ribozymes of the invention may be prepared by any method known in the art for the synthesis of nucleic acid molecules. These include techniques for chemically synthesizing oligonucleotides such as solid phase phosphoramidite chemical synthesis. Alternatively, RNA molecules may be generated by in vitro and in vivo transcription of DNA sequences encoding XMAD. Such DNA sequences may be incorporated into a wide variety of vectors with suitable RNA polymerase promoters such as T7 or SP6. Alternatively, these cDNA constructs that synthesize complementary RNA, constitutively or inducibly, can be introduced into cell lines, cells, or tissues.

[0207] RNA molecules may be modified to increase intracellular stability and half-life. Possible modifications include, but are not limited to, the addition of flanking sequences at the 5′ and/or 3′ ends of the molecule, or the use of phosphorothioate or 2′ O-methyl rather than phosphodiesterase linkages within the backbone of the molecule. This concept is inherent in the production of PNAs and can be extended in all of these molecules by the inclusion of nontraditional bases such as inosine, queosine, and wybutosine, as well as acetyl-, methyl-, thio-, and similarly modified forms of adenine, cytdine, guanine, thymine, and uridine which are not as easily recognized by endogenous endonucleases.

[0208] An additional embodiment of the invention encompasses a method for screening for a compound which is effective in altering expression of a polynucleotide encoding XMAD. Compounds which may be effective in altering expression of a specific polynucleotide may include, but are not limited to, oligonucleotides, antisense oligonucleotides, triple helix-forming oligonucleotides, transcription factors and other polypeptide transcriptional regulators, and non-macromolecular chemical entities which are capable of interacting with specific polynucleotide sequences. Effective compounds may alter polynucleotide expression by acting as either inhibitors or promoters of polynucleotide expression. Thus, in the treatment of disorders associated with increased XMAD expression or activity, a compound which specifically inhibits expression of the polynucleotide encoding XMAD may be therapeutically useful, and in the treament of disorders associated with decreased XMAD expression or activity, a compound which specifically promotes expression of the polynucleotide encoding XMAD may be therapeutically useful.

[0209] At least one, and up to a plurality, of test compounds may be screened for effectiveness in altering expression of a specific polynucleotide. A test compound may be obtained by any method commonly known in the art, including chemical modification of a compound known to be effective in altering polynucleotide expression; selection from an existing, commercially-available or proprietary library of naturally-occurring or non-natural chemical compounds; rational design of a compound based on chemical and/or structural properties of the target polynucleotide; and selection from a library of chemical compounds created combinatorially or randomly. A sample comprising a polynucleotide encoding XMAD is exposed to at least one test compound thus obtained. The sample may comprise, for example, an intact or permeabilized cell, or an in vitro cell-free or reconstituted biochemical system. Alterations in the expression of a polynucleotide encoding XMAD are assayed by any method commonly known in the art. Typically, the expression of a specific nucleotide is detected by hybridization with a probe having a nucleotide sequence complementary to the sequence of the polynucleotide encoding XMAD. The amount of hybridization may be quantified, thus forming the basis for a comparison of the expression of the polynucleotide both with and without exposure to one or more test compounds. Detection of a change in the expression of a polynucleotide exposed to a test compound indicates that the test compound is effective in altering the expression of the polynucleotide. A screen for a compound effective in altering expression of a specific polynucleotide can be carried out, for example, using a Schizosaccharomvces pombe gene expression system (Atkins, D. et al. (1999) U.S. Pat. No. 5,932,435; Arndt, G. M. et al. (2000) Nucleic Acids Res. 28:E15) or a human cell line such as HeLa cell (Clarke, M. L. et al. (2000) Biochem. Biophys. Res. Commun. 268:8-13). A particular embodiment of the present invention involves screening a combinatorial library of oligonucleotides (such as deoxyribonucleotides, ribonucleotides, peptide nucleic acids, and modified oligonucleotides) for antisense activity against a specific polynucleotide sequence (Bruice, T. W. et al. (1997) U.S. Pat. No. 5,686,242; Bruice, T. W. et al. (2000) U.S. Pat. No. 6,022,691).

[0210] Many methods for introducing vectors into cells or tissues are available and equally suitable for use in vivo, in vitro, and ex vivo. For ex vivo therapy, vectors may be introduced into stem cells taken from the patient and clonally propagated for autologous transplant back into that same patient. Delivery by transfection, by liposome injections, or by polycationic amino polymers may be achieved using methods which are well known in the art. (See, e.g., Goldman, C. K. et al. (1997) Nat. Biotechnol. 15:462-466.)

[0211] Any of the therapeutic methods described above may be applied to any subject in need of such therapy, including, for example, mammals such as humans, dogs, cats, cows, horses, rabbits, and monkeys.

[0212] An additional embodiment of the invention relates to the administration of a composition which generally comprises an active ingredient formulated with a pharmaceutically acceptable excipient. Excipients may include, for example, sugars, starches, celluloses, gums, and proteins. Various formulations are commonly known and are thoroughly discussed in the latest edition of Remington's Pharmaceutical Sciences (Maack Publishing, Easton Pa.). Such compositions may consist of XMAD, antibodies to XMAD, and mimetics, agonists, antagonists, or inhibitors of XMAD.

[0213] The compositions utilized in this invention may be administered by any number of routes including, but not limited to, oral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, intraventricular, pulmonary, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, or rectal means.

[0214] Compositions for pulmonary administration may be prepared in liquid or dry powder form. These compositions are generally aerosolized immediately prior to inhalation by the patient. In the case of small molecules (e.g. traditional low molecular weight organic drugs), aerosol delivery of fast-acting formulations is well-known in the art. In the case of macromolecules (e.g. larger peptides and proteins), recent developments in the field of pulmonary delivery via the alveolar region of the lung have enabled the practical delivery of drugs such as insulin to blood circulation (see, e.g., Patton, J. S. et al., U.S. Pat. No. 5,997,848). Pulmonary delivery has the advantage of administration without needle injection, and obviates the need for potentially toxic penetration enhancers.

[0215] Compositions suitable for use in the invention include compositions wherein the active ingredients are contained in an effective amount to achieve the intended purpose. The determination of an effective dose is well within the capability of those skilled in the art.

[0216] Specialized forms of compositions may be prepared for direct intracellular delivery of macromolecules comprising XMAD or fragments thereof. For example, liposome preparations containing a cell-impermeable macromolecule may promote cell fusion and intracellular delivery of the macromolecule. Alternatively, XMAD or a fragment thereof may be joined to a short cationic N-terminal portion from the HIV Tat-l protein. Fusion proteins thus generated have been found to transduce into the cells of all tissues, including the brain, in a mouse model system (Schwarze, S. R. et al. (1999) Science 285:1569-1572).

[0217] For any compound, the therapeutically effective dose can be estimated initially either in cell culture assays, e.g., of neoplastic cells, or in animal models such as mice, rats, rabbits, dogs, monkeys, or pigs. An animal model may also be used to determine the appropriate concentration range and route of administration. Such information can then be used to determine useful doses and routes for administration in humans.

[0218] A therapeutically effective dose refers to that amount of active ingredient, for example XMAD or fragments thereof, antibodies of XMAD, and agonists, antagonists or inhibitors of XMAD, which ameliorates the symptoms or condition. Therapeutic efficacy and toxicity may be determined by standard pharmaceutical procedures in cell cultures or with experimental animals, such as by calculating the ED₅₀ (the dose therapeutically effective in 50% of the population) or LD₅₀ (the dose lethal to 50% of the population) statistics. The dose ratio of toxic to therapeutic effects is the therapeutic index, which can be expressed as the LD₅₀/ED₅₀ ratio. Compositions which exhibit large therapeutic indices are preferred. The data obtained from cell culture assays and animal studies are used to formulate a range of dosage for human use. The dosage contained in such compositions is preferably within a range of circulating concentrations that includes the ED₅₀ with little or no toxicity. The dosage varies within this range depending upon the dosage form employed, the sensitivity of the patient, and the route of administration.

[0219] The exact dosage will be determined by the practitioner, in light of factors related to the subject requiring treatment. Dosage and administration are adjusted to provide sufficient levels of the active moiety or to maintain the desired effect. Factors which may be taken into account include the severity of the disease state, the general health of the subject, the age, weight, and gender of the subject, time and frequency of administration, drug combination(s), reaction sensitivities, and response to therapy. Long-acting compositions may be administered every 3 to 4 days, every week, or biweekly depending on the half-life and clearance rate of the particular formulation.

[0220] Normal dosage amounts may vary from about 0.1 μg to 100,000 μg, up to a total dose of about 1 gram, depending upon the route of administration. Guidance as to particular dosages and methods of delivery is provided in the literature and generally available to practitioners in the art. Those skilled in the art will employ different formulations for nucleotides than for proteins or their inhibitors. Similarly, delivery of polynucleotides or polypeptides will be specific to particular cells, conditions, locations, etc.

[0221] Diagnostics

[0222] In another embodiment, antibodies which specifically bind XMAD may be used for the diagnosis of disorders characterized by expression of XMAD, or in assays to monitor patients being treated with XMAD or agonists, antagonists, or inhibitors of XMAD. Antibodies useful for diagnostic purposes may be prepared in the same manner as described above for therapeutics. Diagnostic assays for XMAD include methods which utilize the antibody and a label to detect XMAD in human body fluids or in extracts of cells or tissues. The antibodies may be used with or without modification, and may be labeled by covalent or non-covalent attachment of a reporter molecule. A wide variety of reporter molecules, several of which are described above, are known in the art and may be used.

[0223] A variety of protocols for measuring XMAD, including ELISAs, RIAs, and FACS, are known in the art and provide a basis for diagnosing altered or abnormal levels of XMAD expression. Normal or standard values for XMAD expression are established by combining body fluids or cell extracts taken from normal mammalian subjects, for example, human subjects, with antibody to XMAD under conditions suitable for complex formation. The amount of standard complex formation may be quantitated by various methods, such as photometric means. Quantities of XMAD expressed in subject, control, and disease samples from biopsied tissues are compared with the standard values. Deviation between standard and subject values establishes the parameters for diagnosing disease.

[0224] In another embodiment of the invention, the polynucleotides encoding XMAD may be used for diagnostic purposes. The polynucleotides which may be used include oligonucleotide sequences, complementary RNA and DNA molecules, and PNAs. The polynucleotides may be used to detect and quantify gene expression in biopsied tissues in which expression of XMAD may be correlated with disease. The diagnostic assay may be used to determine absence, presence, and excess expression of XMAD, and to monitor regulation of XMAD levels during therapeutic intervention.

[0225] In one aspect, hybridization with PCR probes which are capable of detecting polynucleotide sequences, including genomic sequences, encoding XMAD or closely related molecules may be used to identify nucleic acid sequences which encode XMAD. The specificity of the probe, whether it is made from a highly specific region, e.g., the 5′ regulatory region, or from a less specific region, e.g., a conserved motif, and the stringency of the hybridization or amplification will determine whether the probe identifies only naturally occurring sequences encoding XMAD, allelic variants, or related sequences.

[0226] Probes may also be used for the detection of related sequences, and may have at least 50% sequence identity to any of the XMAD encoding sequences. The hybridization probes of the subject invention may be DNA or RNA and may be derived from the sequence of SEQ ID NO:22-42 or from genomic sequences including promoters, enhancers, and introns of the XMAD gene.

[0227] Means for producing specific hybridization probes for DNAs encoding XMAD include the cloning of polynucleotide sequences encoding XMAD or XMAD derivatives into vectors for the production of mRNA probes. Such vectors are known in the art, are commercially available, and may be used to synthesize RNA probes in vitro by means of the addition of the appropriate RNA polymerases and the appropriate labeled nucleotides. Hybridization probes may be labeled by a variety of reporter groups, for example, by radionuclides such as ³²P or ³⁵S, or by enzymatic labels, such as alkaline phosphatase coupled to the probe via avidin/biotin coupling systems, and the like.

[0228] Polynucleotide sequences encoding XMAD may be used for the diagnosis of disorders associated with expression of XMAD. Examples of such disorders include, but are not limited to, a genetic disorder, such as adrenoleukodystrophy, Alport's syndrome, choroideremia, Duchenne and Becker muscular dystrophy, Down's syndrome, cystic fibrosis, chronic granulomatous disease, Gaucher's disease, Huntington's chorea, Marfan's syndrome, muscular dystrophy, myotonic dystrophy, pycnodysostosis, Refsum's syndrome, retinoblastoma, sickle cell anemia, thalassemia, Werner syndrome, von Willebrand's disease, Wilms' tumor, Zellweger syndrome, peroxisomal acyl-CoA oxidase deficiency, peroxisomal thiolase deficiency, peroxisomal bifunctional protein deficiency, mitochondrial carnitine palmitoyl transferase and carnitine deficiency, mitochondrial very-long-chain acyl-CoA dehydrogenase deficiency, mitochondrial medium-chain acyl-CoA dehydrogenase deficiency, mitochondrial short-chain acyl-CoA dehydrogenase deficiency, mitochondrial electron transport flavoprotein and electron transport flavoprotein:ubiquinone oxidoreductase deficiency, mitochondrial trifunctional protein deficiency, and mitochondrial short-chain 3-hydroxyacyl-CoA dehydrogenase deficiency; an autoimmune/inflammation disorder, such as acquired immunodeficiency syndrome (AIDS), Addison's disease, adult respiratory distress syndrome, allergies, ankylosing spondylitis, amyloidosis, anemia, asthma, atherosclerosis, autoimmune hemolytic anemia, autoimmune thyroiditis, autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED), bronchitis, cholecystitis, contact dermatitis, Crohn's disease, atopic dermatitis, dermatomyositis, diabetes mellitus, emphysema, episodic lymphopenia with lymphocytotoxins, erythroblastosis fetalis, erythema nodosum, atrophic gastritis, glomerulonephritis, Goodpasture's syndrome, gout, Graves' disease, Hashimoto's thyroiditis, hypereosinophilia, irritable bowel syndrome, multiple sclerosis, myasthenia gravis, myocardial or pericardial inflammation, osteoarthritis, osteoporosis, pancreatitis, polymyositis, psoriasis, Reiter's syndrome, rheumatoid arthritis, scleroderma, Sjogren's syndrome, systemic anaphylaxis, systemic lupus erythematosus, systemic sclerosis, thrombocytopenic purpura, ulcerative colitis, uveitis, Werner syndrome, complications of cancer, hemodialysis, and extracorporeal circulation, viral, bacterial, fungal, parasitic, protozoal, and helminthic infections, and trauma; and a cell proliferative disorder, such as actinic keratosis, arteriosclerosis, atherosclerosis, bursitis, cirrhosis, hepatitis, mixed connective tissue disease (MCTD), myelofibrosis, paroxysmal nocturnal hemoglobinuria, polycythemia vera, psoriasis, primary thrombocythemia, and cancers including adenocarcinoma, leukemia, lymphoma, melanoma, myeloma, sarcoma, teratocarcinoma, and, in particular, cancers of the adrenal gland, bladder, bone, bone marrow, brain, breast, cervix, gall bladder, ganglia, gastrointestinal tract, heart, kidney, liver, lung, muscle, ovary, pancreas, parathyroid, penis, prostate, salivary glands, skin, spleen, testis, thymus, thyroid, and uterus. The polynucleotide sequences encoding XMAD may be used in Southern or northern analysis, dot blot, or other membrane-based technologies; in PCR technologies; in dipstick, pin, and multiformat ELISA-like assays; and in microarrays utilizing fluids or tissues from patients to detect altered XMAD expression. Such qualitative or quantitative methods are well known in the art.

[0229] In a particular aspect, the nucleotide sequences encoding XMAD may be useful in assays that detect the presence of associated disorders, particularly those mentioned above. The nucleotide sequences encoding XMAD may be labeled by standard methods and added to a fluid or tissue sample from a patient under conditions suitable for the formation of hybridization complexes. After a suitable incubation period, the sample is washed and the signal is quantified and compared with a standard value. If the amount of signal in the patient sample is significantly altered in comparison to a control sample then the presence of altered levels of nucleotide sequences encoding XMAD in the sample indicates the presence of the associated disorder. Such assays may also be used to evaluate the efficacy of a particular therapeutic treatment regimen in animal studies, in clinical trials, or to monitor the treatment of an individual patient.

[0230] In order to provide a basis for the diagnosis of a disorder associated with expression of XMAD, a normal or standard profile for expression is established. This may be accomplished by combining body fluids or cell extracts taken from normal subjects, either animal or human, with a sequence, or a fragment thereof, encoding XMAD, under conditions suitable for hybridization or amplification. Standard hybridization may be quantified by comparing the values obtained from normal subjects with values from an experiment in which a known amount of a substantially purified polynucleotide is used. Standard values obtained in this manner may be compared with values obtained from samples from patients who are symptomatic for a disorder. Deviation from standard values is used to establish the presence of a disorder.

[0231] Once the presence of a disorder is established and a treatment protocol is initiated, hybridization assays may be repeated on a regular basis to determine if the level of expression in the patient begins to approximate that which is observed in the normal subject. The results obtained from successive assays may be used to show the efficacy of treatment over a period ranging from several days to months.

[0232] With respect to cancer, the presence of an abnormal amount of transcript (either under- or overexpressed) in biopsied tissue from an individual may indicate a predisposition for the development of the disease, or may provide a means for detecting the disease prior to the appearance of actual clinical symptoms. A more definitive diagnosis of this type may allow health professionals to employ preventative measures or aggressive treatment earlier thereby preventing the development or further progression of the cancer.

[0233] Additional diagnostic uses for oligonucleotides designed from the sequences encoding XMAD may involve the use of PCR. These oligomers may be chemically synthesized, generated enzymatically, or produced in vitro. Oligomers will preferably contain a fragment of a polynucleotide encoding XMAD, or a fragment of a polynucleotide complementary to the polynucleotide encoding XMAD, and will be employed under optimized conditions for identification of a specific gene or condition. Oligomers may also be employed under less stringent conditions for detection or quantification of closely related DNA or RNA sequences.

[0234] In a particular aspect, oligonucleotide primers derived from the polynucleotide sequences encoding XMAD may be used to detect single nucleotide polymorphisms (SNPs). SNPs are substitutions, insertions and deletions that are a frequent cause of inherited or acquired genetic disease in humans. Methods of SNP detection include, but are not limited to, single-stranded conformation polymorphism (SSCP) and fluorescent SSCP (fSSCP) methods. In SSCP, oligonucleotide primers derived from the polynucleotide sequences encoding XMAD are used to amplify DNA using the polymerase chain reaction (PCR). The DNA may be derived, for example, from diseased or normal tissue, biopsy samples, bodily fluids, and the like. SNPs in the DNA cause differences in the secondary and tertiary structures of PCR products in single-stranded form, and these differences are detectable using gel electrophoresis in non-denaturing gels. In fSCCP, the oligonucleotide primers are fluorescently labeled, which allows detection of the amplimers in high-throughput equipment such as DNA sequencing machines. Additionally, sequence database analysis methods, termed in silico SNP (isSNP), are capable of identifying polymorphisms by comparing the sequence of individual overlapping DNA fragments which assemble into a common consensus sequence. These computer-based methods filter out sequence variations due to laboratory preparation of DNA and sequencing errors using statistical models and automated analyses of DNA sequence chromatograms. In the alternative, SNPs may be detected and characterized by mass spectrometry using, for example, the high throughput MASSARRAY system (Sequenom, Inc., San Diego Calif.).

[0235] Methods which may also be used to quantify the expression of XMAD include radiolabeling or biotinylating nucleotides, coamplification of a control nucleic acid, and interpolating results from standard curves. (See, e.g., Melby, P. C. et al. (1993) J. Immunol. Methods 159:235-244; Duplaa, C. et al. (1993) Anal. Biochem. 212:229-236.) The speed of quantitation of multiple samples may be accelerated by running the assay in a high-throughput format where the oligomer or polynucleotide of interest is presented in various dilutions and a spectrophotometric or colorimetric response gives rapid quantitation.

[0236] In further embodiments, oligonucleotides or longer fragments derived from any of the polynucleotide sequences described herein may be used as elements on a microarray. The microarray can be used in transcript imaging techniques which monitor the relative expression levels of large numbers of genes simultaneously as described in Seilhamer, J. J. et al., “Comparative Gene Transcript Analysis,” U.S. Pat. No. 5,840,484, incorporated herein by reference. The microarray may also be used to identify genetic variants, mutations, and polymorphisms. This information may be used to determine gene function, to understand the genetic basis of a disorder, to diagnose a disorder, to monitor progression/regression of disease as a function of gene expression, and to develop and monitor the activities of therapeutic agents in the treatment of disease. In particular, this information may be used to develop a pharmacogenomic profile of a patient in order to select the most appropriate and effective treatment regimen for that patient. For example, therapeutic agents which are highly effective and display the fewest side effects may be selected for a patient based on his/her pharmacogenomic profile.

[0237] In another embodiment, antibodies specific for XMAD, or XMAD or fragments thereof may be used as elements on a microarray. The microarray may be used to monitor or measure protein-protein interactions, drug-target interactions, and gene expression profiles, as described above.

[0238] A particular embodiment relates to the use of the polynucleotides of the present invention to generate a transcript image of a tissue or cell type. A transcript image represents the global pattern of gene expression by a particular tissue or cell type. Global gene expression patterns are analyzed by quantifying the number of expressed genes and their relative abundance under given conditions and at a given time. (See Seilhamer et al., “Comparative Gene Transcript Analysis,” U.S. Pat. No. 5,840,484, expressly incorporated by reference herein.) Thus a transcript image may be generated by hybridizing the polynucleotides of the present invention or their complements to the totality of transcripts or reverse transcripts of a particular tissue or cell type. In one embodiment, the hybridization takes place in high-throughput format, wherein the polynucleotides of the present invention or their complements comprise a subset of a plurality of elements on a microarray. The resultant transcript image would provide a profile of gene activity.

[0239] Transcript images may be generated using transcripts isolated from tissues, cell lines, biopsies, or other biological samples. The transcript image may thus reflect gene expression in vivo, as in the case of a tissue or biopsy sample, or in vitro, as in the case of a cell line.

[0240] Transcript images which profile the expression of the polynucleotides of the present invention may also be used in conjunction with in vitro model systems and preclinical evaluation of pharmaceuticals, as well as toxicological testing of industrial and naturally-occurring environmental compounds. All compounds induce characteristic gene expression patterns, frequently termed molecular fingerprints or toxicant signatures, which are indicative of mechanisms of action and toxicity (Nuwaysir, E. F. et al. (1999) Mol. Carcinog. 24:153-159; Steiner, S. and N. L. Anderson (2000) Toxicol. Lett. 112-113:467-471, expressly incorporated by reference herein). If a test compound has a signature similar to that of a compound with known toxicity, it is likely to share those toxic properties. These fingerprints or signatures are most useful and refined when they contain expression information from a large number of genes and gene families. Ideally, a genome-wide measurement of expression provides the highest quality signature. Even genes whose expression is not altered by any tested compounds are important as well, as the levels of expression of these genes are used to normalize the rest of the expression data. The normalization procedure is useful for comparison of expression data after treatment with different compounds. While the assignment of gene function to elements of a toxicant signature aids in interpretation of toxicity mechanisms, knowledge of gene function is not necessary for the statistical matching of signatures which leads to prediction of toxicity. (See, for example, Press Release 00-02 from the National Institute of Environmental Health Sciences, released Feb. 29, 2000, available at http://www.niehs.nih.gov/oc/news/toxchip.htm.) Therefore, it is important and desirable in toxicological screening using toxicant signatures to include all expressed gene sequences.

[0241] In one embodiment, the toxicity of a test compound is assessed by treating a biological sample containing nucleic acids with the test compound. Nucleic acids that are expressed in the treated biological sample are hybridized with one or more probes specific to the polynucleotides of the present invention, so that transcript levels corresponding to the polynucleotides of the present invention may be quantified. The transcript levels in the treated biological sample are compared with levels in an untreated biological sample. Differences in the transcript levels between the two samples are indicative of a toxic response caused by the test compound in the treated sample.

[0242] Another particular embodiment relates to the use of the polypeptide sequences of the present invention to analyze the proteome of a tissue or cell type. The term proteome refers to the global pattern of protein expression in a particular tissue or cell type. Each protein component of a proteome can be subjected individually to further analysis. Proteome expression patterns, or profiles, are analyzed by quantifying the number of expressed proteins and their relative abundance under given conditions and at a given time. A profile of a cell's proteome may thus be generated by separating and analyzing the polypeptides of a particular tissue or cell type. In one embodiment, the separation is achieved using two-dimensional gel electrophoresis, in which proteins from a sample are separated by isoelectric focusing in the first dimension, and then according to molecular weight by sodium dodecyl sulfate slab gel electrophoresis in the second dimension (Steiner and Anderson, supra). The proteins are visualized in the gel as discrete and uniquely positioned spots, typically by staining the gel with an agent such as Coomassie Blue or silver or fluorescent stains. The optical density of each protein spot is generally proportional to the level of the protein in the sample. The optical densities of equivalently positioned protein spots from different samples, for example, from biological samples either treated or untreated with a test compound or therapeutic agent, are compared to identify any changes in protein spot density related to the treatment. The proteins in the spots are partially sequenced using, for example, standard methods employing chemical or enzymatic cleavage followed by mass spectrometry. The identity of the protein in a spot may be determined by comparing its partial sequence, preferably of at least 5 contiguous amino acid residues, to the polypeptide sequences of the present invention. In some cases, further sequence data may be obtained for definitive protein identification.

[0243] A proteomic profile may also be generated using antibodies specific for XMAD to quantify the levels of XMAD expression. In one embodiment, the antibodies are used as elements on a microarray, and protein expression levels are quantified by exposing the microarray to the sample and detecting the levels of protein bound to each array element (Lueking, A. et al. (1999) Anal. Biochem. 270:103-111; Mendoze, L. G. et al. (1999) Biotechniques 27:778-788). Detection may be performed by a variety of methods known in the art, for example, by reacting the proteins in the sample with a thiol- or amino-reactive fluorescent compound and detecting the amount of fluorescence bound at each array element.

[0244] Toxicant signatures at the proteome level are also useful for toxicological screening, and should bc analyzed in parallel with toxicant signatures at the transcript level. There is a poor correlation between transcript and protein abundances for some proteins in some tissues (Anderson, N. L. and J. Seilhamer (1997) Electrophoresis 18:533-537), so proteome toxicant signatures may be useful in the analysis of compounds which do not significantly affect the transcript image, but which alter the proteomic profile. In addition, the analysis of transcripts in body fluids is difficult, due to rapid degradation of mRNA, so proteomic profiling may be more reliable and informative in such cases.

[0245] In another embodiment, the toxicity of a test compound is assessed by treating a biological sample containing proteins with the test compound. Proteins that are expressed in the treated biological sample are separated so that the amount of each protein can be quantified. The amount of each protein is compared to the amount of the corresponding protein in an untreated biological sample. A difference in the amount of protein between the two samples is indicative of a toxic response to the test compound in the treated sample. Individual proteins are identified by sequencing the amino acid residues of the individual proteins and comparing these partial sequences to the polypeptides of the present invention.

[0246] In another embodiment, the toxicity of a test compound is assessed by treating a biological sample containing proteins with the test compound. Proteins from the biological sample are incubated with antibodies specific to the polypeptides of the present invention. The amount of protein recognized by the antibodies is quantified. The amount of protein in the treated biological sample is compared with the amount in an untreated biological sample. A difference in the amount of protein between the two samples is indicative of a toxic response to the test compound in the treated sample.

[0247] Microarrays may be prepared, used, and analyzed using methods known in the art. (See, e.g., Brennan, T. M. et al. (1995) U.S. Pat. No. 5,474,796; Schena, M. et al. (1996) Proc. Natl. Acad. Sci. USA 93:10614-10619; Baldeschweiler et al. (1995) PCT application WO95/251116; Shalon, D. et al. (1995) PCT application WO95/35505; Heller, R. A. et al. (1997) Proc. Natl. Acad. Sci. USA 94:2150-2155; and Heller, M. J. et al. (1997) U.S. Pat. No. 5,605,662.) Various types of microarrays are well known and thoroughly described in DNA Microarrays: A Practical Approach, M. Schena, ed. (1999) Oxford University Press, London, hereby expressly incorporated by reference.

[0248] In another embodiment of the invention, nucleic acid sequences encoding XMAD may be used to generate hybridization probes useful in mapping the naturally occurring genomic sequence. Either coding or noncoding sequences may be used, and in some instances, noncoding sequences may be preferable over coding sequences. For example, conservation of a coding sequence among members of a multi-gene family may potentially cause undesired cross hybridization during chromosomal mapping. The sequences may be mapped to a particular chromosome, to a specific region of a chromosome, or to artificial chromosome constructions, e.g., human artificial chromosomes (HACs), yeast artificial chromosomes (YACs), bacterial artificial chromosomes (BACs), bacterial PI constructions, or single chromosome cDNA libraries. (See, e.g., Harrington, J. J. et al. (1997) Nat. Genet. 15:345-355; Price, C.M. (1993) Blood Rev. 7:127-134; and Trask, B. J. (1991) Trends Genet. 7:149-154.) Once mapped, the nucleic acid sequences of the invention may be used to develop genetic linkage maps, for example, which correlate the inheritance of a disease state with the inheritance of a particular chromosome region or restriction fragment length polymorphism (RFLP). (See, e.g., Lander, E. S. and D. Botstein (1986) Proc. Natl. Acad. Sci. USA 83:7353-7357.)

[0249] Fluorescent in situ hybridization (FISH) may be correlated with other physical and genetic map data. (See, e.g., Heinz-Ulrich, et al. (1995) in Meyers, supra, pp. 965-968.) Examples of genetic map data can be found in various scientific journals or at the Online Mendelian Inheritance in Man (OMIM) World Wide Web site. Correlation between the location of the gene encoding XMAD on a physical map and a specific disorder, or a predisposition to a specific disorder, may help define the region of DNA associated with that disorder and thus may further positional cloning efforts.

[0250] In situ hybridization of chromosomal preparations and physical mapping techniques, such as linkage analysis using established chromosomal markers, may be used for extending genetic maps. Often the placement of a gene on the chromosome of another mammalian species, such as mouse, may reveal associated markers even if the exact chromosomal locus is not known. This information is valuable to investigators searching for disease genes using positional cloning or other gene discovery techniques. Once the gene or genes responsible for a disease or syndrome have been crudely localized by genetic linkage to a particular genomic region, e.g., ataxia-telangiectasia to 11q22-23, any sequences mapping to that area may represent associated or regulatory genes for further investigation. (See, e.g., Gatti, R. A. et al. (1988) Nature 336:577-580.) The nucleotide sequence of the instant invention may also be used to detect differences in the chromosomal location due to translocation, inversion, etc., among normal, carrier, or affected individuals.

[0251] In another embodiment of the invention, XMAD, its catalytic or immunogenic fragments, or oligopeptides thereof can be used for screening libraries of compounds in any of a variety of drug screening techniques. The fragment employed in such screening may be free in solution, affixed to a solid support, borne on a cell surface, or located intracellularly. The formation of binding complexes between XMAD and the agent being tested may be measured.

[0252] Another technique for drug screening provides for high throughput screening of compounds having suitable binding affinity to the protein of interest. (See, e.g., Geysen, et al. (1984) PCT application WO84/03564.) In this method, large numbers of different small test compounds are synthesized on a solid substrate. The test compounds are reacted with XMAD, or fragments thereof, and washed. Bound XMAD is then detected by methods well known in the art. Purified XMAD can also be coated directly onto plates for use in the aforementioned drug screening techniques. Alternatively, non-neutralizing antibodies can be used to capture the peptide and immobilize it on a solid support.

[0253] In another embodiment, one may use competitive drug screening assays in which neutralizing antibodies capable of binding XMAD specifically compete with a test compound for binding XMAD. In this manner, antibodies can be used to detect the presence of any peptide which shares one or more antigenic determinants with XMAD.

[0254] In additional embodiments, the nucleotide sequences which encode XMAD may be used in any molecular biology techniques that have yet to be developed, provided the new techniques rely on properties of nucleotide sequences that are currently known, including, but not limited to, such properties as the triplet genetic code and specific base pair interactions.

[0255] Without further elaboration, it is believed that one skilled in the art can, using the preceding description, utilize the present invention to its fullest extent. The following preferred specific embodiments are, therefore, to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever.

[0256] Without further elaboration, it is believed that one skilled in the art can, using the preceding description, utilize the present invention to its fullest extent. The following preferred specific embodiments are, therefore, to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever.

[0257] The disclosures of all patents, applications, and publications mentioned above and below, in particular U.S. Ser. No. 60/172,354, and U.S. Ser. No. 60/172,852, are hereby expressly incorporated by reference.

EXAMPLES

[0258] 1. Construction of cDNA Libraries

[0259] RNA was purchased from Clontech or isolated from tissues described in Table 4. Some tissues were homogenized and lysed in guanidinium isothiocyanate, while others were homogenized and lysed in phenol or in a suitable mixture of denaturants, such as TRIZOL (Life Technologies), a monophasic solution of phenol and guanidine isothiocyanate. The resulting lysates were centrifuged over CsCl cushions or extracted with chloroform. RNA was precipitated from the lysates with either isopropanol or sodium acetate and ethanol, or by other routine methods.

[0260] Phenol extraction and precipitation of RNA were repeated as necessary to increase RNA purity. In some cases, RNA was treated with DNase. For most libraries, poly(A+) RNA was isolated using oligo d(T)-coupled paramagnetic particles (Promega), OLIGOTEX latex particles (QIAGEN, Chatsworth Calif.), or an OLIGOTEX mRNA purification kit (QIAGEN). Alternatively, RNA was isolated directly from tissue lysates using other RNA isolation kits, e.g., the POLY(A)PURE mRNA purification kit (Ambion, Austin Tex.).

[0261] In some cases, Stratagene was provided with RNA and constructed the corresponding cDNA libraries. Otherwise, cDNA was synthesized and cDNA libraries were constructed with the UNIZAP vector system (Stratagene) or SUPERSCRIPT plasmid system (Life Technologies), using the recommended procedures or similar methods known in the art. (See, e.g., Ausubel, 1997, supra, units 5.1-6.6.) Reverse transcription was initiated using oligo d(T) or random primers. Synthetic oligonucleotide adapters were ligated to double stranded cDNA, and the cDNA was digested with the appropriate restriction enzyme or enzymes. For most libraries, the cDNA was size-selected (300-1000 bp) using SEPHACRYL S1000, SEPHAROSE CL2B, or SEPHAROSE CL4B column chromatography (Amersham Pharmacia Biotech) or preparative agarose gel electrophoresis. cDNAs were ligated into compatible restriction enzyme sites of the polylinker of a suitable plasmid, e.g., PBLUESCRIPT plasmid (Stratagene), PSPORT1 plasmid (Life Technologies), pcDNA2.1 plasmid (Invitrogen, Carlsbad Calif.), or pINCY plasmid (Incyte Genomics, Palo Alto Calif.). Recombinant plasmids were transformed into competent E. coli cells including XL1-Blue, XL1-BlueMRF, or SOLR from Stratagene or DH5α, DH10B, or ElectroMAX DH10B from Life Technologies.

[0262] II. Isolation of cDNA Clones

[0263] Plasmids obtained as described in Example I were recovered from host cells by in vivo excision using the UNIZAP vector system (Stratagene) or by cell lysis. Plasmids were purified using at least one of the following: a Magic or WIZARD Minipreps DNA purification system (Promega); an AGTC Miniprep purification kit (Edge Biosystems, Gaithersburg Md.); and QIAWELL 8 Plasmid, QIAWELL 8 Plus Plasmid, QIAWELL 8 Ultra Plasmid purification systems or the R.E.A.L. PREP 96 plasmid purification kit from QIAGEN. Following precipitation, plasmids were resuspended in 0.1 ml of distilled water and stored, with or without lyophilization, at 4° C.

[0264] Alternatively, plasmid DNA was amplified from host cell lysates using direct link PCR in a high-throughput format (Rao, V. B. (1994) Anal. Biochem. 216:1-14). Host cell lysis and thermal cycling steps were carried out in a single reaction mixture. Samples were processed and stored in 384-well plates, and the concentration of amplified plasmid DNA was quantified fluorometrically using PICOGREEN dye (Molecular Probes, Eugene Oreg.) and a FLUOROSKAN II fluorescence scanner (Labsystems Oy, Helsinki, Finland).

[0265] III. Sequencing and Analysis

[0266] Incyte cDNA recovered in plasmids as described in Example II were sequenced as follows. Sequencing reactions were processed using standard methods or high-throughput instrumentation such as the ABI CATALYST 800 (Applied Biosystems) thermal cycler or the PTC-200 thermal cycler (MJ Research) in conjunction with the HYDRA microdispenser (Robbins Scientific) or the MICROLAB 2200 (Hamilton) liquid transfer system. cDNA sequencing reactions were prepared using reagents provided by Amersham Pharmacia Biotech or supplied in ABI sequencing kits such as the ABI PRISM BIGDYE Terminator cycle sequencing ready reaction kit (Applied Biosystems). Electrophoretic separation of cDNA sequencing reactions and detection of labeled polynucleotides were carried out using the MEGABACE 1000 DNA sequencing system (Molecular Dynamics); the ABI PRISM 373 or 377 sequencing system (Applied Biosystems) in conjunction with standard ABI protocols and base calling software; or other sequence analysis systems known in the art. Reading frames within the cDNA sequences were identified using standard methods (reviewed in Ausubel, 1997, supra, unit 7.7). Some of the cDNA sequences were selected for extension using the techniques disclosed in Example VI.

[0267] The polynucleotide sequences derived from cDNA sequencing were assembled and analyzed using a combination of software programs which utilize algorithms well known to those skilled in the art. Table 5 summarizes the tools, programs, and algorithms used and provides applicable descriptions, references, and threshold parameters. The first column of Table 5 shows the tools, programs, and algorithms used, the second column provides brief descriptions thereof, the third column presents appropriate references, all of which are incorporated by reference herein in their entirety, and the fourth column presents, where applicable, the scores, probability values, and other parameters used to evaluate the strength of a match between two sequences (the higher the score, the greater the homology between two sequences). Sequences were analyzed using MACDNASIS PRO software (Hitachi Software Engineering, South San Francisco Calif.) and LASERGENE software (DNASTAR). Polynucleotide and polypeptide sequence alignments were generated using the default parameters specified by the clustal algorithm as incorporated into the MEGALIGN multisequence alignment program (DNASTAR), which also calculates the percent identity between aligned sequences.

[0268] The polynucleotide sequences were validated by removing vector, linker, and polyA sequences and by masking ambiguous bases, using algorithms and programs based on BLAST, dynamic programing, and dinucleotide nearest neighbor analysis. The sequences were then queried against a selection of public databases such as the GenBank primate, rodent, mammalian, vertebrate, and eukaryote databases, and BLOCKS, PRINTS, DOMO, PRODOM, and PFAM to acquire annotation using programs based on BLAST, FASTA, and BLIMPS. The sequences were assembled into full length polynucleotide sequences using programs based on Phred, Phrap, and Consed, and were screened for open reading frames using programs based on GeneMark, BLAST, and FASTA. The full length polynucleotide sequences were translated to derive the corresponding full length amino acid sequences, and these full length sequences were subsequently analyzed by querying against databases such as the GenBank databases (described above), SwissProt, BLOCKS, PRINTS, DOMO, PRODOM, Prosite, and Hidden Markov Model (HMM)-based protein family databases such as PFAM. HMM is a probabilistic approach which analyzes consensus primary structures of gene families. (See, e.g., Eddy, S.R. (1996) Curr. Opin. Struct. Biol. 6:361-365.)

[0269] The programs described above for the assembly and analysis of full length polynucleotide and amino acid sequences were also used to identify polynucleotide sequence fragments from SEQ ID NO:22-42. Fragments from about 20 to about 4000 nucleotides which are useful in hybridization and amplification technologies were described in The Invention section above.

[0270] IV. Analysis of Polynucleotide Expression

[0271] Northern analysis is a laboratory technique used to detect the presence of a transcript of a gene and involves the hybridization of a labeled nucleotide sequence to a membrane on which RNAs from a particular cell type or tissue have been bound. (See, e.g., Sambrook, supra, ch. 7; Ausubel, 1995, supra, ch. 4 and 16.)

[0272] Analogous computer techniques applying BLAST were used to search for identical or related molecules in cDNA databases such as GenBank or LIFESEQ (Incyte Genomics). This analysis is much faster than multiple membrane-based hybridizations. In addition, the sensitivity of the computer search can be modified to determine whether any particular match is categorized as exact or similar. The basis of the search is the product score, which is defined as: $\frac{\text{BLAST~~Score} \times \text{Percent~~Identity}}{5 \times {minimum}\quad \left\{ {{{length}\left( {{Seq}.\quad 1} \right)},{{length}\left( {{Seq}.\quad 2} \right)}} \right\}}$

[0273] The product score takes into account both the degree of similarity between two sequences and the length of the sequence match. The product score is a normalized value between 0 and 100, and is calculated as follows: the BLAST score is multiplied by the percent nucleotide identity and the product is divided by (5 times the length of the shorter of the two sequences). The BLAST score is calculated by assigning a score of +5 for every base that matches in a high-scoring segment pair (HSP), and 4 for every mismatch. Two sequences may share more than one HSP (separated by gaps). If there is more than one HSP, then the pair with the highest BLAST score is used to calculate the product score. The product score represents a balance between fractional overlap and quality in a BLAST alignment. For example, a product score of 100 is produced only for 100% identity over the entire length of the shorter of the two sequences being compared. A product score of 70 is produced either by 100% identity and 70% overlap at one end, or by 88% identity and 100% overlap at the other. A product score of 50 is produced either by 100% identity and 50% overlap at one end, or 79% identity and 100% overlap.

[0274] The results of northern analyses are reported as a percentage distribution of libraries in which the transcript encoding XMAD occurred. Analysis involved the categorization of cDNA libraries by organ/tissue and disease. The organitissue categories included cardiovascular, dermatologic, developmental, endocrine, gastrointestinal, hematopoietic/immune, musculoskeletal, nervous, reproductive, and urologic. The disease/condition categories included cancer, inflammation, trauma, cell proliferation, neurological, and pooled. For each category, the number of libraries expressing the sequence of interest was counted and divided by the total number of libraries across all categories. Percentage values of tissue-specific and disease- or condition-specific expression are reported in Table 3.

[0275] V. Chromosomal Mapping of XMAD Encoding Polynucleotides

[0276] The cDNA sequences which were used to assemble SEQ ID NO:22-42 were compared with sequences from the Incyte LIFESEQ database and public domain databases using BLAST and other implementations of the Smith-Waterman algorithm. Sequences from these databases that matched SEQ ID NO:22-42 were assembled into clusters of contiguous and overlapping sequences using assembly algorithms such as Phrap (Table 5). Radiation hybrid and genetic mapping data available from public resources such as the Stanford Human Genome Center (SHGC), Whitehead Institute for Genome Research (WIGR), and Généthon were used to determine if any of the clustered sequences had been previously mapped. Inclusion of a mapped sequence in a cluster resulted in the assignment of all sequences of that cluster, including its particular SEQ ID NO:, to that map location.

[0277] The genetic map location of SEQ ID NO:18 is described in The Invention as a range, or interval, of a human chromosome. The map position of an interval, in centiMorgans, is measured relative to the terminus of the chromosome's p-arm. (The centiMorgan (cM) is a unit of measurement based on recombination frequencies between chromosomal markers. On average, 1 cM is roughly equivalent to 1 megabase (Mb) of DNA in humans, although this can vary widely due to hot and cold spots of recombination.) The cM distances are based on genetic markers mapped by Généthon which provide boundaries for radiation hybrid markers whose sequences were included in each of the clusters. Human genome maps and other resources available to the public, such as the NCBI “GeneMap'99” World Wide Web site (http://www.ncbi.nlm.nih.gov/genemap/), can be employed to determine if previously identified disease genes map within or in proximity to the intervals indicated above.

[0278] VI. Extension of XMAD Encoding Polynucleotides

[0279] The full length nucleic acid sequences of SEQ ID NO:22-42 were produced by extension of an appropriate fragment of the full length molecule using oligonucleotide primers designed from this fragment. One primer was synthesized to initiate 5′ extension of the known fragment, and the other primer, to initiate 3′ extension of the known fragment. The initial primers were designed using OLIGO 4.06 software (National Biosciences), or another appropriate program, to be about 22 to 30 nucleotides in length, to have a GC content of about 50% or more, and to anneal to the target sequence at temperatures of about 68° C. to about 72° C. Any stretch of nucleotides which would result in hairpin structure and primer-primer dimerizations was avoided.

[0280] Selected human cDNA libraries were used to extend the sequence. If more than one extension was necessary or desired, additional or nested sets of primers were designed.

[0281] High fidelity amplification was obtained by PCR using methods well known in the art. PCR was performed in 96-well plates using the PTC-200 thermal cycler (MJ Research, Inc.). The reaction mix contained DNA template, 200 mmol of each primer, reaction buffer containing Mg²⁺, (NH₄)₂SO₄, and β-mercaptoethanol, Taq DNA polymerase (Amersham Pharmacia Biotech), ELONGASE enzyme (Life Technologies), and Pfu DNA polymerase (Stratagene), with the following parameters for primer pair PCI A and PCI B: Step 1: 94° C., 3 min; Step 2: 94° C., 15 sec; Step 3: 60° C., 1 min; Step 4: 68° C., 2 min; Step 5: Steps 2, 3, and 4 repeated 20 times; Step 6: 68° C., 5 min; Step 7: storage at 4° C. In the alternative, the parameters for primer pair T7 and SK+ were as follows: Step 1: 94° C., 3 min; Step 2: 94° C., 15 sec; Step 3: 57° C., 1 min; Step 4: 68° C., 2 min; Step 5: Steps 2, 3, and 4 repeated 20 times;

[0282] Step 6: 68° C., 5 min; Step 7: storage at 4° C.

[0283] The concentration of DNA in each well was determined by dispensing 100 μl PICOGREEN quantitation reagent (0.25% (v/v) PICOGREEN; Molecular Probes, Eugene Oreg.) dissolved in 1× TE and 0.5 μl of undiluted PCR product into each well of an opaque fluorimeter plate (Corning Costar, Acton Mass.), allowing the DNA to bind to the reagent. The plate was scanned in a Fluoroskan II (Labsystems Oy, Helsinki, Finland) to measure the fluorescence of the sample and to quantify the concentration of DNA. A 5 μl to 10 μl aliquot of the reaction mixture was analyzed by electrophoresis on a 1% agarose mini-gel to determine which reactions were successful in extending the sequence.

[0284] The extended nucleotides were desalted and concentrated, transferred to 384-well plates, digested with CviJI cholera virus endonuclease (Molecular Biology Research, Madison Wis.), and sonicated or sheared prior to religation into pUC 18 vector (Amersham Pharmacia Biotech). For shotgun sequencing, the digested nucleotides were separated on low concentration (0.6 to 0.8%) agarose gels, fragments were excised, and agar digested with Agar ACE (Promega). Extended clones were religated using T4 ligase (New England Biolabs, Beverly Mass.) into pUC 18 vector (Amersham Pharmacia Biotech), treated with Pfu DNA polymerase (Stratagene) to fill-in restriction site overhangs, and transfected into competent E. coli cells. Transformed cells were selected on antibiotic-containing media, and individual colonies were picked and cultured overnight at 37° C. in 384-well plates in LB/2× carb liquid media, The cells were lysed, and DNA was amplified by PCR using Taq DNA polymerase (Amersham Pharmacia Biotech) and Pfu DNA polymerase (Stratagene) with the following parameters: Step 1: 94° C., 3 min; Step 2: 94° C., 15 sec; Step 3: 60° C., 1 min; Step 4: 72° C., 2 min; Step 5: steps 2, 3, and 4 repeated 29 times; Step 6: 72° C., 5 min; Step 7: storage at 4° C. DNA was quantified by PICOGREEN reagent (Molecular Probes) as described above. Samples with low DNA recoveries were reamplified using the same conditions as described above. Samples were diluted with 20% dimethysulfoxide (1;2, v/v), and sequenced using DYENAMIC energy transfer sequencing primers and the DYENAMIC DIRECT kit (Amersham Pharmacia Biotech) or the ABI PRISM BIGDYE Terminator cycle sequencing ready reaction kit (Applied Biosystems).

[0285] In like manner, the polynucleotide sequences of SEQ ID NO:22-42 are used to obtain 5′ regulatory sequences using the procedure above, along with oligonucleotides designed for such extension, and an appropriate genomic library.

[0286] VII. Labeling and Use of Individual Hybridization Probes

[0287] Hybridization probes derived from SEQ ID NO:22-42 are employed to screen cDNAs, genomic DNAs, or mRNAs. Although the labeling of oligonucleotides, consisting of about 20 base pairs, is specifically described, essentially the same procedure is used with larger nucleotide fragments. Oligonucleotides are designed using state-of-the-art software such as OLIGO 4.06 software (National Biosciences) and labeled by combining 50 pmol of each oligomer, 250 μCi of [γ-³²P] adenosine triphosphate (Amersham Pharmacia Biotech), and T4 polynucleotide kinase (DuPont NEN, Boston Mass.). The labeled oligonucleotides are substantially purified using a SEPHADEX G-25 superfine size exclusion dextran bead column (Amersham Pharmacia Biotech). An aliquot containing 107 counts per minute of the labeled probe is used in a typical membrane-based hybridization analysis of human genomic DNA digested with one of the following endonucleases: Ase I, Bgl II, Eco RI, Pst I, Xba I, or Pvu II (DuPont NEN).

[0288] The DNA from each digest is fractionated on a 0.7% agarose gel and transferred to nylon membranes (Nytran Plus, Schlcicher & Schuell, Durham NH). Hybridization is carried out for 16 hours at 40° C. To remove nonspecific signals, blots are sequentially washed at room temperature under conditions of up to, for example, 0× saline sodium citrate and 0.5% sodium dodecyl sulfate. Hybridization patterns are visualized using autoradiography or an alternative imaging means and compared.

[0289] VIII. Microarrays

[0290] The linkage or synthesis of array elements upon a microarray can be achieved utilizing photolithography, piezoelectric printing (ink-jet printing, See, e.g., Baldeschweiler, supra), mechanical microspotting technologies, and derivatives thereof. The substrate in each of the aforementioned technologies should be uniform and solid with a non-porous surface (Schena (1999), supra). Suggested substrates include silicon, silica, glass slides, glass chips, and silicon wafers. Alternatively, a procedure analogous to a dot or slot blot may also be used to arrange and link elements to the surface of a substrate using thermal, UV, chemical, or mechanical bonding procedures. A typical array may be produced using available methods and machines well known to those of ordinary skill in the art and may contain any appropriate number of elements. (See, e.g., Schena, M. et al. (1995) Science 270:467-470; Shalon, D. et al. (1996) Genome Res. 6:639-645; Marshall, A. and J. Hodgson (1998) Nat. Biotechnol. 16:27-31.)

[0291] Full length cDNAs, Expressed Sequence Tags (ESTs), or fragments or oligomers thereof may comprise the elements of the microarray. Fragments or oligomers suitable for hybridization can be selected using software well known in the art such as LASERGENE software (DNASTAR). The array elements are hybridized with polynucleotides in a biological sample. The polynucleotides in the biological sample are conjugated to a fluorescent label or other molecular tag for ease of detection. After hybridization, nonhybridized nucleotides from the biological sample are removed, and a fluorescence scanner is used to detect hybridization at each array element. Alternatively, laser desorbtion and mass spectrometry may be used for detection of hybridization. The degree of complementarity and the relative abundance of each polynucleotide which hybridizes to an element on the microarray may be assessed. In one embodiment, microarray preparation and usage is described in detail below.

[0292] Tissue or Cell Sample Preparation

[0293] Total RNA is isolated from tissue samples using the guanidinium thiocyanate method and poly(A)⁺ RNA is purified using the oligo-(dT) cellulose method. Each poly(A)⁺ RNA sample is reverse transcribed using MMLV reverse-transcriptase, 0.05 pg/μl oligo-(dT) primer (21 mer), 1× first strand buffer, 0.03 units/μl RNase inhibitor, 500 μM dATP, 500 μM dGTP, 500 μM dTTP, 40 μM dCTP, 40 μM dCTP-Cy3 (BDS) or dCTP-Cy5 (Amersham Pharmacia Biotech). The reverse transcription reaction is performed in a 25 ml volume containing 200 ng poly(A)⁺ RNA with GEMBRIGHT kits (Incyte). Specific control poly(A)⁺ RNAs are synthesized by in vitro transcription from non-coding yeast genomic DNA. After incubation at 37° C. for 2 hr, each reaction sample (one with Cy3 and another with Cy5 labeling) is treated with 2.5 ml of 0.5M sodium hydroxide and incubated for 20 minutes at 85° C. to the stop the reaction and degrade the RNA. Samples are purified using two successive CHROMA SPIN 30 gel filtration spin columns (CLONTECH Laboratories, Inc. (CLONTECH), Palo Alto Calif.) and after combining, both reaction samples are ethanol precipitated using 1 ml of glycogen (1 mg/ml), 60 ml sodium acetate, and 300 ml of 100% ethanol. The sample is then dried to completion using a SpeedVAC (Savant Instruments Inc., Holbrook N.Y.) and resuspended in 14 μl 5× SSC/0.2% SDS.

[0294] Microarray Preparation

[0295] Sequences of the present invention are used to generate array elements. Each array element is amplified from bacterial cells containing vectors with cloned cDNA inserts. PCR amplification uses primers complementary to the vector sequences flanking the cDNA insert. Array elements are amplified in thirty cycles of PCR from an initial quantity of 1-2 ng to a final quantity greater than 5 μg. Amplified array elements are then purified using SEPHACRYL-400 (Amersham Pharmacia Biotech).

[0296] Purified array elements are immobilized on polymer-coated glass slides. Glass microscope slides (Corning) are cleaned by ultrasound in 0.1% SDS and acetone, with extensive distilled water washes between and after treatments. Glass slides are etched in 4% hydrofluoric acid (VWR Scientific Products Corporation (VWR), West Chester Pa.), washed extensively in distilled water, and coated with 0.05% aminopropyl silane (Sigma) in 95% ethanol. Coated slides are cured in a 110° C. oven.

[0297] Array elements are applied to the coated glass substrate using a procedure described in U.S. Pat. No. 5,807,522, incorporated herein by reference. 1 μl of the array element DNA, at an average concentration of 100 ng/μl, is loaded into the open capillary printing element by a high-speed robotic apparatus. The apparatus then deposits about 5 nl of array element sample per slide.

[0298] Microarrays are UV-crosslinked using a STRATALINKER UV-crosslinker (Stratagene). Microarrays are washed at room temperature once in 0.2% SDS and three times in distilled water. Non-specific binding sites are blocked by incubation of microarrays in 0.2% casein in phosphate buffered saline (PBS) (Tropix, Inc., Bedford Mass.) for 30 minutes at 60° C. followed by washes in 0.2% SDS and distilled water as before.

[0299] Hybridization

[0300] Hybridization reactions contain 9 μl of sample mixture consisting of 0.2 μg each of Cy3 and Cy5 labeled cDNA synthesis products in 5× SSC, 0.2% SDS hybridization buffer. The sample mixture is heated to 65° C. for 5 minutes and is aliquoted onto the microarray surface and covered with an 1.8 cm² coverslip. The arrays are transferred to a waterproof chamber having a cavity just slightly larger than a microscope slide. The chamber is kept at 100% humidity internally by the addition of 140 μl of 5× SSC in a corner of the chamber. The chamber containing the arrays is incubated for about 6.5 hours at 60° C. The arrays are washed for 10 min at 45° C. in a first wash buffer (1× SSC, 0.1% SDS), three times for 10 minutes each at 45° C. in a second wash buffer (0.1× SSC), and dried.

[0301] Detection

[0302] Reporter-labeled hybridization complexes are detected with a microscope equipped with an Innova 70 mixed gas 10 W laser (Coherent, Inc., Santa Clara Calif.) capable of generating spectral lines at 488 nm for excitation of Cy3 and at 632 nm for excitation of Cy5. The excitation laser light is focused on the array using a 20× microscope objective (Nikon, Inc., Melville N.Y.). The slide containing the array is placed on a computer-controlled X-Y stage on the microscope and raster-scanned past the objective. The 1.8 cm×1.8 cm array used in the present example is scanned with a resolution of 20 micrometers.

[0303] In two separate scans, a mixed gas multiline laser excites the two fluorophores sequentially. Emitted light is split, based on wavelength, into two photomultiplier tube detectors (PMT R1477, Hamamatsu Photonics Systems, Bridgewater N.J.) corresponding to the two fluorophores. Appropriate filters positioned between the array and the photomultiplier tubes are used to filter the signals. The emission maxima of the fluorophores used are 565 nm for Cy3 and 650 nm for Cy5. Each array is typically scanned twice, one scan per fluorophore using the appropriate filters at the laser source, although the apparatus is capable of recording the spectra from both fluorophores simultaneously.

[0304] The sensitivity of the scans is typically calibrated using the signal intensity generated by a cDNA control species added to the sample mixture at a known concentration. A specific location on the array contains a complementary DNA sequence, allowing the intensity of the signal at that location to be correlated with a weight ratio of hybridizing species of 1:100,000. When two samples from different sources (e.g., representing test and control cells), each labeled with a different fluorophore, are hybridized to a single array for the purpose of identifying genes that are differentially expressed, the calibration is done by labeling samples of the calibrating cDNA with the two fluorophores and adding identical amounts of each to the hybridization mixture.

[0305] The output of the photomultiplier tube is digitized using a 12-bit RTI-835H analog-to-digital (A/D) conversion board (Analog Devices, Inc., Norwood Mass.) installed in an IBM-compatible PC computer. The digitized data are displayed as an image where the signal intensity is mapped using a linear 20-color transformation to a pseudocolor scale ranging from blue (low signal) to red (high signal). The data is also analyzed quantitatively. Where two different fluorophores are excited and measured simultaneously, the data are first corrected for optical crosstalk (due to overlapping emission spectra) between the fluorophores using each fluorophore's emission spectrum.

[0306] A grid is superimposed over the fluorescence signal image such that the signal from each spot is centered in each element of the grid. The fluorescence signal within each element is then integrated to obtain a numerical value corresponding to the average intensity of the signal. The software used for signal analysis is the GEMTOOLS gene expression analysis program (Incyte).

[0307] IX. Complementary Polynucleotides

[0308] Sequences complementary to the XMAD-encoding sequences, or any parts thereof, are used to detect, decrease, or inhibit expression of naturally occurring XMAD. Although use of oligonucleotides comprising from about 15 to 30 base pairs is described, essentially the same procedure is used with smaller or with larger sequence fragments. Appropriate oligonucleotides are designed using OLIGO 4.06 software (National Biosciences) and the coding sequence of XMAD. To inhibit transcription, a complementary oligonucleotide is designed from the most unique 5′ sequence and used to prevent promoter binding to the coding sequence. To inhibit translation, a complementary oligonucleotide is designed to prevent ribosomal binding to the XMAD-encoding transcript.

[0309] X. Expression of XMAD

[0310] Expression and purification of XMAD is achieved using bacterial or virus-based expression systems. For expression of XMAD in bacteria, cDNA is subcloned into an appropriate vector containing an antibiotic resistance gene and an inducible promoter that directs high levels of cDNA transcription. Examples of such promoters include, but are not limited to, the trp-lac (tac) hybrid promoter and the T5 or T7 bacteriophage promoter in conjunction with the lac operator regulatory element. Recombinant vectors are transformed into suitable bacterial hosts, e.g., BL21(DE3). Antibiotic resistant bacteria express XMAD upon induction with isopropyl beta-D-thiogalactopyranoside (IPTG). Expression of XMAD in eukaryotic cells is achieved by infecting insect or mammalian cell lines with recombinant Autographica californica nuclear polyhedrosis virus (AcMNPV), commonly known as baculovirus. The nonessential polyhedrin gene of baculovirus is replaced with cDNA encoding XMAD by either homologous recombination or bacterial-mediated transposition involving transfer plasmid intermediates. Viral infectivity is maintained and the strong polyhedrin promoter drives high levels of cDNA transcription. Recombinant baculovirus is used to infect Spodoptera frugiperda (Sf9) insect cells in most cases, or human hepatocytes, in some cases. Infection of the latter requires additional genetic modifications to baculovirus. (See Engelhard, E. K. et al. (1994) Proc. Natl. Acad. Sci. USA 91:3224-3227; Sandig, V. et al. (1996) Hum. Gene Ther. 7:1937-1945.)

[0311] In most expression systems, XMAD is synthesized as a fusion protein with, e.g., glutathione S-transferase (GST) or a peptide epitope tag, such as FLAG or 6-His, permitting rapid, single-step, affinity-based purification of recombinant fusion protein from crude cell lysates. GST, a 26-kilodalton enzyme from Schistosoma japonicum, enables the purification of fusion proteins on immobilized glutathione under conditions that maintain protein activity and antigenicity (Amersham Pharmacia Biotech). Following purification, the GST moiety can be proteolytically cleaved from XMAD at specifically engineered sites. FLAG, an 8-amino acid peptide, enables immunoaffinity purification using commercially available monoclonal and polyclonal anti-FLAG antibodies (Eastman Kodak). 6-His, a stretch of six consecutive histidine residues, enables purification on metal-chelate resins (QIAGEN). Methods for protein expression and purification are discussed in Ausubel (1995, supra, ch. 10 and 16). Purified XMAD obtained by these methods can be used directly in the assays shown in Examples XI and XV.

[0312] XI. Demonstration of XMAD Activity

[0313] An assay for XMAD activity measures the disruption of cytoskeletal filament networks upon overexpression of XMAD in cultured cell lines. (Rezniczek, G. A. et al. (1998) J. Cell Biol. 141:209-225.) cDNA encoding XMAD is subcloned into a mammalian expression vector that drives high levels of cDNA expression. This construct is transfected into cultured cells, such as rat kangaroo PtK2 or rat bladder carcinoma 804G cells. Actin filaments and intermediate filaments such as keratin and vimentin are visualized by immunofluorescence microscopy using antibodies and techniques well known in the art. The configuration and abundance of cyoskeletal filaments can be assessed and quantified using confocal imaging techniques. In particular, the bundling and collapse of cytoskeletal filament networks is indicative of XMAD activity.

[0314] Alternatively, an assay for XMAD activity measures the amount of cell aggregation induced by overexpression of XMAD. In this assay, cultured cells such as NIH3T3 are transfected with cDNA encoding XMAD contained within a suitable mammalian expression vector under control of a strong promoter. Cotransfection with cDNA encoding a fluorescent marker protein, such as Green Fluorescent Protein (Clontech), is useful for identifying stable transfectants. The amount of cell agglutination, or clumping, associated with transfected cells is compared with that associated with untransfected cells. The amount of cell agglutination is a direct measure of XMAD activity.

[0315] Alternatively, protein kinase activity is measured by quantifying the phosphorylation of a protein substrate by XMAD in the presence of gamma-labeled ³²P-ATP. XMAD is incubated with the protein substrate, ³²P-ATP, and an appropriate kinase buffer. The ³²P incorporated into the substrate is separated from free ³²P-ATP by electrophoresis and the incorporated ³²P is counted using a radioisotope counter. The amount of incorporated ³²P is proportional to the activity of XMAD. A determination of the specific amino acid residue phosphorylated is made by phosphoamino acid analysis of the hydrolyzed protein.

[0316] XII. Functional Assays

[0317] XMAD function is assessed by expressing the sequences encoding XMAD at physiologically elevated levels in mammalian cell culture systems. cDNA is subcloned into a mammalian expression vector containing a strong promoter that drives high levels of cDNA expression. Vectors of choice include pCMV SPORT plasmid (Life Technologies) and pCR3.1 plasmid (Invitrogen), both of which contain the cytomegalovirus promoter. 5-10 μg of recombinant vector are transiently transfected into a human cell line, for example, an endothelial or hematopoietic cell line, using either liposome formulations or electroporation. 1-2 μl of an additional plasmid containing sequences encoding a marker protein are co-transfected. Expression of a marker protein provides a means to distinguish transfected cells from nontransfected cells and is a reliable predictor of cDNA expression from the recombinant vector. Marker proteins of choice include, e.g., Green Fluorescent Protein (GFP; Clontech), CD64, or a CD64-GFP fusion protein. Flow cytometry (FCM), an automated, laser optics-based technique, is used to identify transfected cells expressing GFP or CD64-GFP and to evaluate the apoptotic state of the cells and other cellular properties. FCM detects and quantifies the uptake of fluorescent molecules that diagnose events preceding or coincident with cell death. These events include changes in nuclear DNA content as measured by staining of DNA with propidium iodide; changes in cell size and granularity as measured by forward light scatter and 90 degree side light scatter; down-regulation of DNA synthesis as measured by decrease in bromodeoxyuridine uptake; alterations in expression of cell surface and intracellular proteins as measured by reactivity with specific antibodies; and alterations in plasma membrane composition as measured by the binding of fluorescein-conjugated Annexin V protein to the cell surface. Methods in flow cytometry are discussed in Ormerod, M. G. (1994) Flow Cytometry, Oxford, New York N.Y.

[0318] The influence of XMAD on gene expression can be assessed using highly purified populations of cells transfected with sequences encoding XMAD and either CD64 or CD64-GFP. CD64 and CD64-GFP are expressed on the surface of transfected cells and bind to conserved regions of human immunoglobulin G (IgG). Transfected cells are efficiently separated from nontransfected cells using magnetic beads coated with either human IgG or antibody against CD64 (DYNAL, Lake Success N.Y.). mRNA can be purified from the cells using methods well known by those of skill in the art. Expression of mRNA encoding XMAD and other genes of interest can be analyzed by northern analysis or microarray techniques.

[0319] XIII. Production of XMAD Specific Antibodies

[0320] XMAD substantially purified using polyacrylamide gel electrophoresis (PAGE; see, e.g., Harrington, M. G. (1990) Methods Enzymol. 182:488-495), or other purification techniques, is used to immunize rabbits and to produce antibodies using standard protocols.

[0321] Alternatively, the XMAD amino acid sequence is analyzed using LASERGENE software (DNASTAR) to determine regions of high immunogenicity, and a corresponding oligopeptide is synthesized and used to raise antibodies by means known to those of skill in the art. Methods for selection of appropriate epitopes, such as those near the C-terminus or in hydrophilic regions are well described in the art. (See, e.g., Ausubel, 1995, supra, ch. 11.)

[0322] Typically, oligopeptides of about 15 residues in length are synthesized using an ABI 431A peptide synthesizer (Applied Biosystems) using FMOC chemistry and coupled to KLH (Sigma-Aldrich, St. Louis Mo.) by reaction with N-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS) to increase immunogenicity. (See, e.g., Ausubel, 1995, supra.) Rabbits are immunized with the oligopeptide-KLH complex in complete Freund's adjuvant. Resulting antisera are tested for antipeptide and anti-XMAD activity by, for example, binding the peptide or XMAD to a substrate, blocking with 1% BSA, reacting with rabbit antisera, washing, and reacting with radio-iodinated goat anti-rabbit IgG.

[0323] XIV. Purification of Naturally Occurring XMAD Using Specific Antibodies

[0324] Naturally occurring or recombinant XMAD is substantially purified by immunoaffinity chromatography using antibodies specific for XMAD. An immunoaffinity column is constructed by covalently coupling anti-XMAD antibody to an activated chromatographic resin, such as CNBr-activated SEPHAROSE (Amersham Pharmacia Biotech). After the coupling, the resin is blocked and washed according to the manufacturer's instructions.

[0325] Media containing XMAD are passed over the immunoaffinity column, and the column is washed under conditions that allow the preferential absorbance of XMAD (e.g., high ionic strength buffers in the presence of detergent). The column is eluted under conditions that disrupt antibody/XMAD binding (e.g., a buffer of pH 2 to pH 3, or a high concentration of a chaotrope, such as urea or thiocyanate ion), and XMAD is collected.

[0326] XV. Identification of Molecules Which Interact with XMAD

[0327] XMAD, or biologically active fragments thereof, are labeled with ¹²⁵I Bolton-Hunter reagent. (See, e.g., Bolton A. E. and W. M. Hunter (1973) Biochem. J. 133:529-539.) Candidate molecules previously arrayed in the wells of a multi-well plate are incubated with the labeled XMAD, washed, and any wells with labeled XMAD complex are assayed. Data obtained using different concentrations of XMAD are used to calculate values for the number, affinity, and association of XMAD with the candidate molecules.

[0328] Alternatively, molecules interacting with XMAD are analyzed using the yeast two-hybrid system as described in Fields, S. and O. Song (1989, Nature 340:245-246), or using commercially available kits based on the two-hybrid system, such as the MATCHMAKER system (Clontech).

[0329] XMAD may also be used in the PATHCALLING process (CuraGen Corp., New Haven Conn.) which employs the yeast two-hybrid system in a high-throughput manner to determine all interactions between the proteins encoded by two large libraries of genes (Nandabalan, K. et al. (2000) U.S. Pat. No. 6,057,101).

[0330] Various modifications and variations of the described methods and systems of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with certain embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention which are obvious to those skilled in molecular biology or related fields are intended to be within the scope of the following claims. Protein SEQ ID Nucleotide NO: SEQ ID NO: Clone ID Library Fragments  1 22 1424691 BEPINON01 998379R1 (KIDNTUT01), 1424691H1 (BEPINON01), SXAE02538V1  2 23 1450801 PENITUT01 046316H1 (CORNNOT01), 1450801CT1 (PENITUT01), 1450801H1 (PENITUT01), 1671961H1 (BLADNOT05)  3 24 1597872 BRAINOT14 814997R1 (OVARTUT01), 814997T1 (OVARTUT01), 1412857T6 (BRAINOT12), 1438406F1 (PANCNOT08), 1597872H1 (BRAINOT14), 1797683H1 (PORSTUT05), 3346671H1 (BRAITUT24)  4 25 1674661 BLADNOT05 1655227F6 (PROSTUT08), 1674661H1 (BLADNOT05), 1675239F6 (BLADNOT05), 1879940F6 (LEUKNOT03), 2121172F6 (BRSTNOT07), 2157008F6 (BRAINOT09), 2672389F6 (KIDNNOT19), 3270393H1 (BRAINOT20), 3387668H1 (LUNGTUT17), 3685486H1 (HEAANOT01), 4103531H1 (BRSTTUT17), 4850546H1 (TESTNOT10), 5028429H1 (COLCDIT01), 5661414H1 (BRAUNOT01)  5 26 1689337 PROSTUT10 542204R1 (OVARNOT02), 961047R2 (BRSTTUT03), 1623395T6 (BRAITUT13), 1663607F6 (BRSTNOT09), 1689337H1 (PROSTUT10), 2898863R6 (THYMNON04), 3507526H1 (CONCNOT01)  6 27 1746392 STOMTUT02 682990H1 (UTRSNOT02), 1663009F6 (BRSTNOT09), 1746392H1 (STOMTUT02), 1746392T6 (STOMTUT02), 2079257F6 (ISLTNOT01), 3099537F6 (PTHYNOT03), 3111943H1 (BRSTNOT17), 3391682H1 (LUNGNOT28), 4747243F6 (SMCRUNT01)  7 28 1825182 LSUBNOT03 983441H1 (TONGTUT01), 1825182F6 (LSUBNOT03), 1825182H1 (LSUBNOT03), 1825369F6 (LSUBNOT03), SAQB00477F1, SAQB00879F1, SAQB01310F1, SAQB00187F1, SAQA02142F1, SAQA00159F1  8 29 2155541 BRAINOT09 871127T1 (LUNGAST01), 1309342R1 (COLNFET02), 1544021T1 (PROSTUT04), 2155541H1 (BRAINOT09), 2155541X15F1 (BRAINOT09)  9 30 2215706 SINTFET03 570718H1 (MMLR3DT01), 756160R1 (BRAITUT02), 1511501F1 (LUNGNOT14), 2215706F6 (SINTFET03), 2215706H1 (SINTFET03), 2648753F6 (OVARNOT10), 2804428H1 (PENCNOT01), 3092672T6 (BRSTNOT19), 3597972H1 (FIBPNOT01), 3604953H1 (LUNGNOT30), 3879505H1 (SPLNNOT11), 4506625F6 (OVARTDT01), 4708188H1 (BRAIFET02), 4985942H1 (LIVRTUT10), 5151981H1 (HEARFET03), 5644791H1 (UTRSTMR01), 5862219H1 (BRAYDIT01) 10 31 2347692 TESTTUT02 075856R1 (THP1PEB01), 370791R1 (LUNGNOT02), 1502478F1 (BRAITUT07), 2347692H1 (TESTTUT02), 2825041F6 (ADRETUT06) 11 32 2579048 KIDNTUT13 841019R1 (PROSTUT05), 1352253F1 (LATRTUT02), 1414589F6 (BRAINOT12), 1427648F1 (SINTBST01), 1996374R6 (BRSTTUT03), 2579048H1 (KIDNTUT13) 12 33 2604493 LUNGTUT07 901679X18 (BRSTTUT03), 927970X54R1 (BRAINOT04), 1435427F6 (PANCNOT08), 1484806F6 (CORPNOT02), 1962694T6 (BRSTNOT04), 1990921F6 (CORPNOT02), 2279985R6 (PROSNON01), 2279985T6 (PROSNON01), 2294223T6 (BRAINON01), 2604493H1 (LUNGTUT07), 2707717H1 (PONSAZT01), 3421936H1 (UCMCNOT04), 4769752H1 (BRATNOT02), 4989101H1 (LIVRTUT10), SAEA01968R1 13 34 2787182 BRSTNOT13 916228H1 (BRSTNOT04) 1251624F6 (LUNGFET03), 1440454F6 (THYRNOT03), 1664091F6 (BRSTNOT09), 1812788X17C1 (PROSTUT12), 1812788X21C1 (PROSTUT12), 2787182H1 (BRSTNOT13), 2824324F6 (ADRETUT06), 2882479T6 (UTRSTUT05), 3833543H1 (PANCNOT17), 60147041D2, 60147042B6, 60147044D2, SXAE05916V1, SXAE02927V1, g1670173 14 35 3096668 CERVNOT03 2373962F6 (ISLTNOT01), 2373962T6 (ISLTNOT01), 2762988H1 (BRSTNOT12) , 3096668F6 (CERVNOT03), 3096668H1 (CERVNOTO3) , 3096668T6 (CERVNOT03), SCGA06156V1, SCGA11275V1, SCGA07741V1 15 36 3143411 HNT2AZS07 540532T6 (LNODNOT02), 852710R1 (NGANNOT01), 860567R1 (BRAITUT03), 1402106F6 (LATRTUT02), 3143411H1 (HNT2AZS07), 3143411R6 (HNT2AZS07), 5135819H1 (OVARDIT04) 16 37 3170835 BRSTNOT18 3170835H1 (BRSTNOT18), 3171275F6 (BRSTNOT18) 17 38 3550808 SYNONOT01 00101F1 (U937NOT01), 1353706T1 (LATRTUT02), 1426227F1 (SINTBST01), 1804230F6 (SINTNOT13), 2361183T6 (LUNGFET05), 2606392H1 (LUNGTUT07), 3550808H1 (SYNONOT01), SBAA00101F1 18 39 3683905 HEAANOT01 833556H1 (PROSNOT07), 1494051H1 (PROSNON01), 3683905H1 (HEAANOT01), 5512558H1 (BRADDIR01), 5700822H1 (DRGCNOT01), g1267581 19 40 4062841 BRAINOT21 1863239H1 (PROSNOT19), 1863239T6 (PROSNOT19), 4062841H1 (BRAINOT21) 20 41 6394358 UTRENOT10 875733R6 (LUNGAST01), 1312637T6 (BLADTUT02), 2296386R6 (BRSTNOT05), 2296386T6 (BRSTNOT05), 6394358H1 (UTRENOT10) 21 42 2847752 g4126329.v113.gs_6.edit.5p 1-6416; g3449297 5802-10044 5547763H1 (TESTNOC01) 8322-8532; 3373379H1 (CONNTUT05) 8722-8984; 3331371H1 (BRAIFET01) 9378-9641; 5376974H1 (BRAXNOT01) 9634-9772; 4015537F6 (BRAXNOT01) 9709-10226; 5921447H1 (BRAIFET02) 9963-10235; 4700084F6 (BRALNOT01) 10218-10739; 670937H1 (CRBLNOT01) 10517-10780; 3788576H1 (BRAHNOT05) 10608-10904; 5929111H1 (BRAIFET02) 10791- 11069; 3278762T6 (STOMFET02) 11029-11627; 2847752R6 (HNT2AZS07) 11181-11648

[0331] TABLE 2 Amino Potential Potential Analytical Polypeptide Acid Phosphorylation Glycosylation Homologous Methods and SEQ ID NO: Residues Sites Sites Signature Sequence Sequence Databases  1 (1424691) 222 S25 T88 S155 S179 N7 N53 N68 M1-G29: Signal g7649266 MOTIFS T191 T207 S90 S96 peptide Sterile-alpha BLAST_GENBANK S113 S119 E173-E181: 5- motif and SPSCAN hydroxytryptamine leucine zipper BLIMPS-PRINTS 5A (serotonin) containing receptor kinase AZK R213-D215: Rgd cell interaction motif  2 (1450801) 228 S177 S99 N25 F20-G211: Leucine g3786312 MOTIFS Rich Repeat Extracellular BLAST_GENBANK matrix protein HMMER PFAM (Nishiu, J. et BLIMPS-PRINTS al. Genomics (1998) 52:378- 381)  3 (1597872) 386 S88 S137 S229 G237-P247: Insulin- g4033606 MOTIFS S364 T370 like growth factor Extension BLAST_GENBANK G202-R341: (Kieliszewski BLIMPS-BLOCKS Spliceosome- M.J., Lamport BLAST-DOMO associated protein D.T. Plant J. (1994) 5:157- 172)  4 (1674661) 833 S46 T276 T418 S34 N106 N121 M1-V23: Signal g1110599 MOTIFS T57 T229 T302 N310 N419 peptide Semaphorin BLAST_GENBANK S382 T429 S505 N522 N564 F53-K481: homolog SIGPEPT S826 S200 S364 Semaphorin domain (Ignagaki; S. SPSCAN S480 T523 S555 et al. FEBS BLAST-PFAM T561 S685 T701 Lett (1995) S742 Y249 Y345 370:269-272) Y736  5 (1689337) 410 T358 S394 S139 N3 M1-A16: Signal g3450883 MOTIFS S249 S17 S343 peptide Fibroin BLAST_GENBANK T385 S401 P240-Q255 (Gosline, J.M. SPSCAN Prokaryotic et al. J. Exp. BLIMPS-BLOCKS molybdopterin Biol. (1999) BLIMPS-PRINTS oxidoreductase 202:3295-3303) L55-A65: Prepro- orexin signature R349-D351: Rgd cell interaction motif  6 (1746392) 360 S217 S255 T344 E17-P356: g310200 MOTIFS S37 S8 T28 S69 Neurofilament proline-rich BLAST_GENBANK S113 T182 S188 triplet H proteoglycan BLAST-DOMO S224 S242 S250  7 (1825182) 377 S18 S41 S140 T267 N97 N128 N135 F20-G211: Leucine g188864 MOTIFS S38 S62 S120 T343 N146 Rich Repeat mucin BLAST_GENBANK (Shimomura, HMMER-PFAM T., Blood BLIMPS-PRINTS (1990) 75:2349-2356)  8 (2155541) 182 T116 M1-F22: Signal g6164953 MOTIFS peptide vacuolar BLAST_GENBANK T42-D62: Neutrophil sorting SPSCAN cytosol factor protein VPS29 BLIMPS-PRINTS D170-E178: 7-fold [Mus musculus] BLIMPS-PFAM repeat proteins I Edgar, A.J. motif and Polak, R60-D62: Rgd cell J.M. (2000) interaction motif Biochem. Biophys. Res. Commun. 277:622-630  9 (2215706) 513 S239 S325 T14 S51 N69 N89 N338 S94-Q108: g4322670 MOTIFS S71 S110 S137 N429 Adrenocorticotrophin Dentin BLIMPS-PRINTS S171 S208 S213 receptor phosphoryn BLIMPS-PFAM T219 S263 S268 S328-S335: “Phage” P = 7.6e − 07 S349 S394 S403 integrase family S404 T96 T118 R196-D198: Rgd cell S149 S239 S365 interaction motif T475 S479 10 (2347692) 361 T81 T53 S158 S257 N8 N51 N306 G242-M268: C1q g1562534 MOTIFS T333 S128 N324 domain proteins csdp single- BLAST_GENBANK L146-P150: Laminin stranded DNA BLIMPS-BLOCKS G domain protein binding BLIMPS-PFAM G110-P288: protein BLAST-DOMO Fibrillar collagen carboxyl-terminus 11 (2579048) 327 S18 S199 T55 T72 N158 K41-T55: Histone H5 MOTIFS S73 T285 S51 S140 signature BLIMPS_PRINTS S177 S262 R213-D215: Rgd cell interaction motif 12 (2604493) 1 110 S213 T795 S8 T17 N168 N472 V532-V565, L577- g1702924 p0071 MOTIFS S40 S64 S82 T118 N640 N671 F598, M609-L622: Catenin- BLAST_GENBANK S132 S151 S170 N672 N691 Armadillo/beta- related HMMER-PFAM S192 S249 T317 N698 N729 catenin-like protein BLIMPS-PFAM T357 T476 S601 N747 N851 repeats (Hatzfeld M., BLAST-PRODOM S642 S673 S674 N966 S511-H1025: Mouse Nachtsheim C. S701 S731 T795 p120 protein J. Cell Sci. S1015 S1059 S1073 R971-D973: Rgd cell (1996) T84 S236 S276 interaction motif 109:2767-2778) T292 T309 S337 S457 T506 T744 S749 S813 T945 S976 T1078 13 (2787182) 386 S123 T210 S265 N14 N173 C57-Q72: g3876060 MOTIFS S283 S317 S326 Transmembrane motif Weak BLAST_GENBANK T330 T338 T44 S79 M279-M363 similarity HMMER S100 S109 T127 osteonectin with nitrogen PROFILESCAN T142 T170 T214 fixation S332 regulator 14 (3096668) 181 S114 S116 S118 N17 F16-E25: Alpha-type g3393011 MOTIFS S120 S122 T124 calcitonin Clumping BLAST_GENBANK S154 T173 T13 signature factor B BLIMPS-PRINTS (Ni Eidhin D. et al. Mol. Microbiol. 1998 30:245- 257) 15 (3143411) 374 S291 S75 S92 T206 N109 N304 M1-G24: Signal g3790610 MOTIFS T214 T298 T315 peptide Layilin BLAST_GENBANK T23 T37 T50 S51 N224-W247, V328- (Borowsky SIGPEPT S262 T263 S306 N348: Transmembrane M.L., Hynes SPSCAN motif R.O. (1998) J. HMMER Y46-C63, W163-C176: Cell Biol. BLAST-PFAM C-type lectin 143:429-442) BLIMPS-PRINTS domain PROFILESCAN BLAST-DOMO 16 (3170835) 102 S45 T57 T44 N33 M1-T19: Signal g2565394 MOTIFS peptide Cuticle 12 BLAST_GENBANK A55-103: Insect SPSCAN cuticle protein BLAST-PFAM PROFILESCAN BLIMPS-PRINTS BLAST-PRODOM BLAST-DOMO 17 (3550808) 510 S76 S88 S128 S150 N72 N136 N193 M1-G20: Signal g294502 MOTIFS T152 T308 T448 N253 N352 peptide Olfactomedin BLAST_GENBANK T461 S49 S56 S110 N411 L5-G23: (Yokoe H., SIGPEPT T138 Transmembrane motif Anholt R.R. SPSCAN G33-F46: Pheromone Proc. Natl. HMMER B alpha-1 receptor Acad. Sci. USA BLIMPS-PRINTS (1993) 90:4665-9) 18 (3683905) 185 S11 S57 S173 T135 M1-L170: von g2654431 MOTIFS Willebrand factor Type XII BLAST_GENBANK domain score collagen HMMER-PFAM M2-F15, R37-F51, BLIMPS-PRINTS V103-G111 BLAST-PRODOM M1-R171: collagen BLAST-DOMO glycoprotein precursor 19 (4062841) 207 T83 S201 T74 S166 V93-T174: PDZ g3885828 MOTIFS T174 S190 domain (Also known Lin-7-A BLAST_GENBANK as DHR of GLGF) (Irie M. et HMMER-PFAM P91-V171: SH3 al. Oncogene BLIMPS-PRINTS domain (1999) BLIMPS-PFAM 18:2811-2817) BLIMPS-PRODOM BLAST-PRODOM BLAST-DOMO 20 (6394358) 238 S2 S96 S100 S12 N160 L3-A25, T64-E110, MOTIFS S26 T149 S200 E209-R218: 7-fold BLIMPS-PFAM T203 repeat proteins (clathrin) R136-D138: Rgd cell interaction motif 21 3298 S93 T307 S2706 N631 N846 L1536-Y1553 g3449288 MEGF2 MOTIFS S29 S222 S240 N1181 N1221 L2525-A2545 [Rattus BLAST_GENBANK T266 T403 T421 N1316 N1326 I2669-L2689 norvegicus] HMMER T440 T457 T533 N1648 N1712 S2712-A2730 SPSCAN T567 T569 S658 N1769 N2034 L2741-N2761: HMMER-PFAM T744 T756 T778 N2163 N2182 Transmembrane BLIMPS-BLOCKS T831 S865 S880 N2372 N2460 Domains PROFILESCAN S882 T931 T944 N2492 N2683 M1-E31: Signal BLIMPS-PRINTS S983 T985 S1030 N2732 N2794 peptide T1089 T1223 T1246 N3240 Y329-A423; Y437- T1304 S1347 S1366 L535 T1433 T1465 T1488 Y549-V641; F655- S1578 S1630 S1685 L746 T1841 S1903 T2184 Y760-T848; Y862- S2359 S2368 T2482 N952 S2513 S2663 S2847 Y965-Q1057; F1071- T2863 S2878 S2880 V1159 S2885 S2897 T2964 Y1178-I1265: T2979 S2980 S3053 Cadherin domains S3281 S186 S194 C1378-C1431; C1438- T200 T250 S409 C1469 S497 T527 S545 C1478-C1512; C1725- S833 T852 T1020 C1756 S1180 S1230 T1275 C1931-C1962; C1966- S1659 S1660 S1737 C2000: EGF domains S1829 S1976 T2066 F1542-D1704. F1792- S2113 T2216 S2322 E1920: S2355 S2549 S2552 Laminin G domains T2709 S2819 S2822 C2110-A2137; C2588- S2830 S2919 S2956 L2613: T2964 T3273 Y399 GPCR signature Y449 Y842 Y2299

[0332] Polynucleotide Tissue Expression Disease or Condition SEQ ID NO: Fragments (Fraction of Total) (Fraction of Total) Vector 22 1-517 Cardiovascular (0.200) Cell proliferation (0.400) pSPORT1 852-905 Gastrointestinal (0.200) Cell proliferation/Cell line (0.200) Urologic (0.222) Inflammation/Trauma (0.300) 23 1-2387 Reproductive (0.397) Cell proliferation (0.559) pINCY Gastrointestinal (0.132) Inflammation/Trauma (0.176) Musculoskeletal (0.118) Other (0.118) 24 1-901 Reproductive (0.262) Cell proliferation (0.536) pINCY 1456-1471 Gastrointestinal (0.179) Inflammation/Trauma (0.297) Nervous (0.179) Cell proliferation/Cell line (0.190) 25 1-1928 Reproductive (0.294) Cell proliferation (0.471) 1776-3293 Gastrointestinal (0.157) Inflammation/Trauma (0.373) Nervous (0.157) Cell proliferation/Cell line (0.118) pINCY 26 1-821 Reproductive (0.351) Cell proliferation (0.486) 1312-1324 Hematopoietic/Immune (0.135) Cell proliferation/Cell line (0.243) Nervous (0.135) Inflammation/Trauma (0.351) 27 1-626 Reproductive (0.227) Cell proliferation (0.432) pINCY 1034-1324 Nervous (0.182) Cell proliferation/Cell line (0.205) Gastrointestinal (0.159) Inflammation/Trauma (0.228) 28 1-2429 Gastrointestinal (1.000) Cell proliferation (1.000) pINCY 29 1-50 Reproductive (0.197) Cell proliferation (0.441) pINCY 591-985 Nervous (0.164) Inflammation/Trauma (0.454) Hematopoietic/Immune (0.145) Cell proliferation/Cell line (0.191) 30 1-285 Nervous (0.235) Cell proliferation (0.445) pINCY 813-930 Reproductive (0.235) Cell proliferation/Cell line (0.202) 1145-3381 Gastrointestinal (0.126) Inflammation/Trauma (0.252) 31 1-82 Reproductive (0.267) Cell proliferation (0.383) pINCY 1728-1803 Nervous (0.233) Inflammation/Trauma (0.383) Other (0.117) Cell proliferation/Cell line (0.217) 32 1-430 Reproductive (0.263) Cell proliferation (0.544) pINCY 752-964 Nervous (0.193) Cell proliferation/cell line (0.158) 1405-1515 Cardiovascular (0.140) Inflammation/Trauma (0.246) 33 1-570 Nervous (0.383) Cell proliferation (0.395) pINCY 860-1573 Reproductive (0.210) Inflammation/Trauma (0.334) 1859-2494 Gastrointestinal (0.111) Cell proliferation/Cell line (0.198) 2864-4416 34 1-30 Reproductive (0.280) Cell proliferation (0.415) pINCY 190-234 Nervous (0.195) Inflammation/Trauma (0.366) 889-4428 Hematopoietic/Immune (0.146) Cell proliferation/Cell line (0.146) 35 1-189 Reproductive (0.333) Cell proliferation (0.333) pINCY 264-1907 Cardiovascular (0.167) Cell proliferation.Cell line (0.333) Developmental (0.167) Inflammation/Trauma (0.375) 36 1-773 Nervous (0.312) Cell proliferation (0.531) pSPORT1 1742-1839 Reproductive (0.312) Inflmmation/Trauma (0.250) Gastrointestinal (0.125) Cell proliferation/Cell line (0.125) 37 1-503 Reproductive (1.000) Cell proliferation (1.000) pINCY 38 1-167 Gastrointestinal (0.723) Cell proliferation (0.447) pINCY 449-946 Reproductive (0.128) Inflammation/Trauma (0.489) 1541-2154 Urologic (0.085) Trauma (0.170) 39 1-431 Reproductive (0.450) Cell proliferation (0.750) pINCY 666-733 Nervous (0.250) Inflamation/Trauma (0.200) Urologic (0.100) Trauma (0.100) 40 1-48 Reproductive (0.300) Cell proliferation (0.400) pINCY 301-453 Cardiovascular (0.200) Cell proliferation/Cell line (0.300) 634-665 Hematopoietic/Immune (0.200) Inflammation/Trauma (0.400) 41 1-276 Hematopoietic/Immune (0.349) Inflammation/Trauma (0.512) pINCY 553-741 Nervous (0.163) Cell proliferation (0.349) 820-1235 Gastrointestinal (0.140) Cell proliferation/Cell line (0.209)

[0333] TABLE 4 Polynucleotide SEQ ID NO: Library Library Comment 22 BEPINON01 This normalized bronchial epithelium library was constructed from 5.12 million independent clones from the BEPINOT01 library. RNA was made from a bronchial epithelium primary cell line derived from a 54-year-old Caucasian male. The normalization and hybridization conditions were adapted from Soares et al., PNAS (1994) 91:9228, using a longer (24-hour) reannealing hybridization period. 23 PENITUT01 Library was constructed using RNA isolated from tumor tissue removed from the penis of a 64-year-old Caucasian male during penile amputation. Pathology indicated a fungating invasive grade 4 squamous cell carcinoma involving the inner wall of the foreskin and extending onto the glans penis. Patient history included benign neoplasm of the large bowel, atherosclerotic coronary artery disease, angina pectoris, gout, and obesity. Family history included maglignant pharyngeal neoplasm, chronic lymphocytic leukemia, and chronic liver disease. 24 BRAINOT14 Library was constructed using RNA isolated from brain tissue removed from the left frontal lobe of a 40-year-old Caucasian female during excision of a cerebral meningeal lesion. Pathology for the associated tumor tissue indicated grade 4 gemistocytic astrocytoma. 25 BLADNOT05 Library was constructed using RNA isolated from bladder tissue removed from a 60- year-old Caucasian male during a radical cystectomy, prostatectomy, and vasectomy. Pathology for the associated tumor tissue indicated grade 3 transitional cell carcinoma. Carcinoma in-situ was identified in the dome and trigone. Patient history included tobacco use. 26 PROSTUT10 Library was constructed using RNA isolated from prostatic tumor tissue removed from a 66-year-old Caucasian male during radical prostatectom and regional lymph node excision. Pathology indicated an adenocarcinoma (Gleason grade 2 + 3). Adenofibromatous hyperplasia was also present. The patient presented with elevated prostate specific antigen (PSA) . Family history included prostate cancer and secondary bone cancer. 27 STOMTUT02 Library was constructed using RNA isolated from stomach tumor tissue obtained from a 68-year-old Caucasian female during a partial gastrectomy. Pathology indicated a malignant lymphoma of diffuse large-cell type. Previous surgeries included cholecystectomy. Patient history included thalassemia. Family history included acute leukemia, malignant neoplasm of the esophagus, malignant stomach neoplasm, and atherosclerotic coronary artery disease. 28 LSUBNOT03 Library was constructed using RNA isolated from submandibular gland tissue obtained from a 68-year-old Caucasian male during a sialoadenectomy. Family history included acute myocardial infarction, atherosclerotic coronary artery disease, and type II diabetes. 29 BRAINOT09 Library was constructed using RNA isolated from brain tissue removed from a Caucasian male fetus, who died at 23 weeks' gestation. 30 SINTFET03 Library was constructed using RNA isolated from small intestine tissue removed from a Caucasian female fetus, who died at 20 weeks' gestation 31 TESTTUT02 Library was constructed using RNA isolated from testicular tumor removed from a 31- year-old Caucasian male during unilateral orchiectomy. Pathology indicated embryonal carcinoma. 32 KIDNTUT13 Library was constructed using RNA isolated from kidney tumor tissue removed from a 51-year-old Caucasian female during a nephroureterectomy. Patholgy indicated a grade 3 renal cell carcinoma. Patient history included depressive disorder, hypoglycemia, and uterine endometriosis. Family history included calculus of the kidney, colon cancer, and type II diabetes. 33 LUNGTUT07 Library was constructed using RNA isolated from lung tumor tissue removed from the upper lobe of a 50-year-old Caucasian male during segmental lung resection. Pathology indicated an invasive grade 4 squamous cell adenocarcinoma. Patient history included tobacco use. Family history included skin cancer 34 BRSTNOT13 Library was constructed using RNA isolated from breast tissue removed from the left medial lateral breast of a 36-year-old Caucasian female during bilateral simple mastectomy and total breast reconstruction. Pathology indicated benign breast tissue. Patient history included a breast neoplasm, depressive disorder, hyperlipidemia, chronic stomach ulcer, and an ectopic pregnancy. Family history included myocardial infarction, cerebrovascular disease, atherosclerotic coronary artery disease, hyperlipidemia, skin cancer, breast cancer, depressive disorder, esophageal cancer, bone cancer, Hodgkin's lymphoma. bladder cancer, and heart condition. 35 CERVNOT03 Library was constructed using RNA isolated from uterine cervical tissue removed from a 40-year-old Caucasian female during a vaginal hysterectomy with bilateral salpingo-oophorectomy and dilation and curettage. Pathology indicated secretory phase endometrium. 36 HNT2AZS07 This subtracted library was constructed from RNA isolated from an hNT2 cell line (derived from a human teratocarcinoma that exhibited properties characteristic of a committed neuronal precursor) treated for three days with 0.35 micromolar AZ. The hybridization probe for subtraction was derived from a similarly constructed library from untreated hNT2 cells. 3.08 M clones from the AZ-treated library were subjected to three rounds of subtractive hybridization with 3.04 M clones from the untreated library. Subtractive hybridization conditions were based on the methodologies of Swaroop et al. (NAR (1991) 19:1954) and Bonaldo et al. (Genome Research (1996) 6:791). 37 BRSTNOT18 Library was constructed using RNA isolated from diseased breast tissue removed from a 57-year-old Caucasian female during a unilateral simple extended mastectomy. Pathology indicated mildly proliferative breast disease. Patient history included breast cancer and osteoarthritis. Family history included type II diabetes, gallbladder and breast cancer, and chronic lymphocytic leukemia. 38 SYNONOT01 Library was constructed using RNA isolated from synovial tissue removed from a 75- year-old Caucasian male. 39 HEAANOT01 Library was constructed using RNA isolated from right coronary and right circumflex coronary artery tissue removed from the explanted heart of a 46-year-old Caucasian male during a heart transplantation. Patient history included myocardial infarction from total occlusion of the left anterior descending coronary artery, atherosclerotic coronary artery disease, hyperlipiderma, myocardial ischemia, dilated cardiomyopathy, left ventricular dysfunction, and tobacco abuse. Previous surgeries included cardiac catheterization. Family history included atherosclerotic coronary artery disease. 40 BRAINOT21 Library was constructed using RNA isolated from diseased brain tissue removed from the left frontal lobe of a 46-year-old Caucasian male during a lobectomy. Pathology indicated focal cortical and subcortical scarring of the left frontal lobe, characterized by cavitation and extensive reactive changes, including marked gliosis and hemosiderin deposition, consistent with a history of remote severe head trauma. GFAP was positive in astrocytes. The pattern of reactivity is that of reactive gliosis. Patient history included traumatic intracranial hemorrhage and brain injury with loss of consciousness following head trauma. Family history included cerebrovascular disease, cerebrovascular disease, and atherosclerotic coronary artery disease. 41 UTRENOT10 Library was constructed using polyA RNA isolated from pooled uterine endometrial tissue removed from three adult females during endometrial biopsy. Pathology indicated normal endometrium. 42 The Incyte cDNSa for SEQ ID No:42 were derived from cDNA libraries constructed from brain, including tissues associated with Huntington's disease, Alzheimer's disease, and multiple sclerosis, as well as from pituitary, testicular, stomach, spinal cord, kidney, and prostate tissues, and from ovarian, cervical, pancreatic, and soft tissue tumors.

[0334] TABLE 5 Program Description Reference Parameter Threshold ABI A program that removes vector sequences and Perkin-Elmer Applied Biosystems, FACTURA masks ambiguous bases in nucleic acid sequences. Foster City, CA. ABI/ A Fast Data Finder useful in comparing Perkin-Elmer Applied Biosystems, Mismatch <50% PARACEL and annotating amino acid or nucleic Foster City, CA; Paracel Inc., Pasadena, CA. FDF acid sequences. ABI A program that assembles nucleic acid Perkin-Elmer Applied Auto- sequences. Biosystems Foster Assembler City, CA. BLAST A Basic Local Alignment Search Tool useful Altschul, S.F. et al. (1990) J. Mol. Biol. qESTs: Probability value = in sequence similarity search for amino acid 215:403-410; Altschul, S.F. et al. (1997) 1.0E−8 or less and nucleic acid sequences. BLAST includes Nucleic Acids Res. 25: 3389-3402 Full length sequences: Probablity five functions: blastp, blastn, blastx, value = 1.0E−10 or less tblastn, and tblastx. FASTA A Pearson and Lipman algorithm that searches for Pearson, W.R. and D.J. Lipman (1988) Proc. ESTs: fasta E value = 1.06E−6 similarity between a query sequence and a group of Natl. Acad Sci. 85:2444-2448; Pearson, W.R. Assembled ESTs: fasta Identity = sequences of the same type. FASTA comprises (1990) Methods Enzymol. 183:63-98; and 95% or greater and as least five functions: fasta, tfasta, Smith, T.F. and M. S. Waterman (1981) Adv. Match length = 200 fastx, tfastx, and ssearch. Appl. Math. 2:482-489. bases or greater; fastx E value = 1.0E−8 or less Full length sequesnces: fastx score = 100 or greater BLIMPS A BLocks IMProved Searcher that matches a Henikoff, S and J.G. Henikoff, Nucl. Acid Score = 1000 sequence against those in BLOCKS, PRINTS, Res., 19:6565-72, 1991. J.G. Henikoff and S. or greater; Ratio of DOMO, PRODOM, and PFAM databases to Henikoff(1996) Methods Enzymol. 266:88- Score/Strength = search for gene families, sequence 105; and Attwood, T.K. et al. (1997) J. Chem. 0.75 or larger; and, homology, and structural fingerprint regions. Inf. Comput. Sci. 37: 417-424. if applicable, Probability value = 1.0E−3 or less HMMER An algorithm for searching a query sequence against Krogh, A. et al. (1994) J. Mol. Biol., Score = 10-50 bits for PFAM hits, hidden Markov model (HMM)-based databases of 235:1501-1531; Sonnhammer, E.L.L. et al. depending on individual protein protein family consensus sequences, such as PFAM. (1988) Nucleic Acids Res. 26:320-322. families ProfileScan An algorithm that searches for structural Gribskov, M. et al. (1988) CABIOS 4:61-66; Normalized quality score ≧ GCG- and sequence motifs in protein sequences that Gribskov, et al. (1989) Methods Enzymol. specified “HIGH” value for that match sequence patterns defined in Prosite. 183:146-159; Bairoch, A. et al. (1997) particular Prosite motif. Nucleic Acids Res. 25: 217-221. Generally, score = 1.4-2.1 Phred A base-calling algorithm that examines automated Ewing, B. et al. (1998) Genome sequencer traces with high sensitivity Res. 8:175-185; Ewing, B. and P. and probability. Green (1998) Genome Res. 8:186-194 Phrap A Phils Revised Assembly Program including Smith, T.F. and M. S. Waterman (1981) Adv. Score = or greater SWAT and CrossMatch, programs based on Appl. Math. 2:482-489; Smith, T.F. amd M. Match length = 56 or greater efficient implementation of the S. Waterman (1981) J. Mol. Biol. 147:195- Smith-Waterman algorithm, useful in searching 197; and Green, P., University of sequence homology and assembling Washington, Seattle, WA. DNA sequences. Consed A graphical tool for viewing and editing Gordon, D. et al. (1998) Genome Phrap assemblies Res. 8:195-202. SPScan A weight matrix analysis program that scans protein Nielson, H. et al. (1997) Protein Engineering Score = 3.5 or greater sequences for the presence of secretory 10:1-6; Claverie, J.M. and S. Audic (1997) signal peptides. CABIOS 12: 431-439. Motifs A program that searches amino acid Bairoch et al. supra; Wisconsin sequences for patterns that matched those Package Program Manual, version defined in Prosite. 9, page M51-59, Genetics Computer Group, Madison, WI.

[0335]

1 42 1 222 PRT Homo sapiens misc_feature Incyte ID No 1424691CD1 1 Met Arg Gln Ile Ala Ser Asn Thr Ser Leu Gln Arg Ser Gln Ser 1 5 10 15 Asn Pro Ile Leu Gly Ser Pro Phe Phe Ser His Phe Asp Gly Gln 20 25 30 Asp Ser Tyr Ala Ala Ala Val Arg Arg Pro Gln Val Pro Ile Lys 35 40 45 Tyr Gln Gln Ile Thr Pro Val Asn Gln Ser Arg Ser Ser Ser Pro 50 55 60 Thr Gln Tyr Gly Leu Thr Lys Asn Phe Ser Ser Leu His Leu Asn 65 70 75 Ser Arg Asp Ser Gly Phe Ser Ser Gly Asn Thr Asp Thr Ser Ser 80 85 90 Glu Arg Gly Arg Tyr Ser Asp Arg Ser Arg Asn Lys Tyr Gly Arg 95 100 105 Gly Ser Ile Ser Leu Asn Ser Ser Pro Arg Gly Arg Tyr Ser Gly 110 115 120 Lys Ser Gln His Ser Thr Pro Ser Arg Gly Arg Tyr Pro Gly Lys 125 130 135 Phe Tyr Arg Val Ser Gln Ser Ala Leu Asn Pro His Gln Ser Pro 140 145 150 Asp Phe Lys Arg Ser Pro Arg Asp Leu His Gln Pro Asn Thr Ile 155 160 165 Pro Gly Met Pro Leu His Pro Glu Thr Asp Ser Arg Ala Ser Glu 170 175 180 Glu Asp Ser Lys Val Ser Glu Gly Gly Trp Thr Lys Val Glu Tyr 185 190 195 Arg Lys Lys Pro His Arg Pro Ser Pro Ala Lys Thr Asn Lys Glu 200 205 210 Arg Ala Arg Gly Asp His Arg Gly Trp Arg Asn Phe 215 220 2 228 PRT Homo sapiens misc_feature Incyte ID No 1450801CD1 2 Met Ile Leu His Asn Gln Ile Thr Gly Ile Gly Arg Glu Asp Phe 1 5 10 15 Ala Thr Thr Tyr Phe Leu Glu Glu Leu Asn Leu Ser Tyr Asn Arg 20 25 30 Ile Thr Ser Pro Gln Val His Arg Asp Ala Phe Arg Lys Leu Arg 35 40 45 Leu Leu Arg Ser Leu Asp Leu Ser Gly Asn Arg Leu His Thr Leu 50 55 60 Pro Pro Gly Leu Pro Arg Asn Val His Val Leu Lys Val Lys Arg 65 70 75 Asn Glu Leu Ala Ala Leu Ala Arg Gly Ala Leu Ala Gly Met Ala 80 85 90 Gln Leu Arg Glu Leu Tyr Leu Thr Ser Asn Arg Leu Arg Ser Arg 95 100 105 Ala Leu Gly Pro Arg Ala Trp Val Asp Leu Ala His Leu Gln Leu 110 115 120 Leu Asp Ile Ala Gly Asn Gln Leu Thr Glu Ile Pro Glu Gly Leu 125 130 135 Pro Glu Ser Leu Glu Tyr Leu Tyr Leu Gln Asn Asn Lys Ile Ser 140 145 150 Ala Val Pro Ala Asn Ala Phe Asp Ser Thr Pro Asn Leu Lys Gly 155 160 165 Ile Phe Leu Arg Phe Asn Lys Leu Ala Val Gly Ser Val Val Asp 170 175 180 Ser Ala Phe Arg Arg Leu Lys His Leu Gln Val Leu Asp Ile Glu 185 190 195 Gly Asn Leu Glu Phe Gly Asp Ile Ser Lys Asp Arg Gly Arg Leu 200 205 210 Gly Lys Glu Lys Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu 215 220 225 Glu Thr Arg 3 386 PRT Homo sapiens misc_feature Incyte ID No 1597872CD1 3 Met Val His Phe Gln Ala Ser Glu Val Gln Gln Leu Leu His Asn 1 5 10 15 Lys Phe Val Val Ile Leu Gly Asp Ser Ile Gln Arg Ala Val Tyr 20 25 30 Lys Asp Leu Val Leu Leu Leu Gln Lys Asp Ser Leu Leu Thr Ala 35 40 45 Ala Gln Leu Lys Ala Lys Tyr Leu Glu Asp Val Leu Glu Glu Leu 50 55 60 Thr Tyr Gly Pro Ala Pro Asp Leu Val Ile Ile Asn Ser Cys Leu 65 70 75 Trp Asp Leu Ser Arg Tyr Gly Arg Cys Ser Met Glu Ser Tyr Arg 80 85 90 Glu Asn Leu Glu Arg Val Phe Val Arg Met Asp Gln Val Leu Pro 95 100 105 Asp Ser Cys Leu Leu Val Trp Asn Met Ala Met Pro Leu Gly Glu 110 115 120 Arg Ile Thr Gly Gly Phe Leu Leu Pro Glu Leu Gln Pro Leu Ala 125 130 135 Gly Ser Leu Arg Arg Asp Val Val Glu Gly Asn Phe Tyr Ser Ala 140 145 150 Thr Leu Ala Gly Asp His Cys Phe Asp Val Leu Asp Leu His Phe 155 160 165 His Phe Arg His Ala Val Gln His Arg His Arg Asp Gly Val His 170 175 180 Trp Asp Gln His Ala His Arg His Leu Ser His Leu Leu Leu Thr 185 190 195 His Val Ala Asp Ala Trp Gly Val Glu Leu Pro Lys Arg Gly Tyr 200 205 210 Pro Pro Asp Pro Trp Ile Glu Asp Trp Ala Glu Met Asn His Pro 215 220 225 Phe Gln Gly Ser His Arg Gln Thr Pro Asp Phe Gly Glu His Leu 230 235 240 Ala Leu Leu Pro Pro Pro Pro Ser Ser Leu Pro Pro Pro Met Pro 245 250 255 Phe Pro Tyr Pro Leu Pro Gln Pro Ser Pro Pro Pro Leu Phe Pro 260 265 270 Pro Leu Pro Gln Asp Thr Pro Phe Phe Pro Gly Gln Pro Phe Pro 275 280 285 Pro His Glu Phe Phe Asn Tyr Asn Pro Val Glu Asp Phe Ser Met 290 295 300 Pro Pro His Leu Gly Cys Gly Pro Gly Val Asn Phe Val Pro Gly 305 310 315 Pro Leu Pro Pro Pro Ile Pro Gly Pro Asn Pro His Gly Gln His 320 325 330 Trp Gly Pro Val Val His Arg Gly Met Pro Arg Tyr Val Pro Asn 335 340 345 Ser Pro Tyr His Val Arg Arg Met Gly Gly Pro Cys Arg Gln Arg 350 355 360 Leu Arg His Ser Glu Arg Leu Ile His Thr Tyr Lys Leu Asp Arg 365 370 375 Arg Pro Pro Ala His Ser Gly Thr Trp Pro Gly 380 385 4 833 PRT Homo sapiens misc_feature Incyte ID No 1674661CD1 4 Met Ala Pro His Trp Ala Val Trp Leu Leu Ala Ala Arg Leu Trp 1 5 10 15 Gly Leu Gly Ile Gly Ala Glu Val Trp Trp Asn Leu Val Pro Arg 20 25 30 Lys Thr Val Ser Ser Gly Glu Leu Ala Thr Val Val Arg Arg Phe 35 40 45 Ser Gln Thr Gly Ile Gln Asp Phe Leu Thr Leu Thr Leu Thr Glu 50 55 60 Pro Thr Gly Leu Leu Tyr Val Gly Ala Arg Glu Ala Leu Phe Ala 65 70 75 Phe Ser Met Glu Ala Leu Glu Leu Gln Gly Ala Ile Ser Trp Glu 80 85 90 Ala Pro Val Glu Lys Lys Thr Glu Cys Ile Gln Lys Gly Lys Asn 95 100 105 Asn Gln Thr Glu Cys Phe Asn Phe Ile Arg Phe Leu Gln Pro Tyr 110 115 120 Asn Ala Ser His Leu Tyr Val Cys Gly Thr Tyr Ala Phe Gln Pro 125 130 135 Lys Cys Thr Tyr Val Asn Met Leu Thr Phe Thr Leu Glu His Gly 140 145 150 Glu Phe Glu Asp Gly Lys Gly Lys Cys Pro Tyr Asp Pro Ala Lys 155 160 165 Gly His Ala Gly Leu Leu Val Asp Gly Glu Leu Tyr Ser Ala Thr 170 175 180 Leu Asn Asn Phe Leu Gly Thr Glu Pro Ile Ile Leu Arg Asn Met 185 190 195 Gly Pro His His Ser Met Lys Thr Glu Tyr Leu Ala Phe Trp Leu 200 205 210 Asn Glu Pro His Phe Val Gly Ser Ala Tyr Val Pro Glu Thr Val 215 220 225 Gly Ser Phe Thr Gly Asp Asp Asp Lys Val Tyr Phe Phe Phe Arg 230 235 240 Glu Arg Ala Leu Glu Ser Asp Cys Tyr Ala Glu Gln Val Val Ala 245 250 255 Arg Val Ala Arg Val Cys Lys Gly Asp Met Gly Gly Ala Arg Thr 260 265 270 Leu Gln Arg Lys Trp Thr Thr Phe Leu Lys Ala Arg Leu Ala Cys 275 280 285 Ser Ala Pro Asn Trp Gln Leu Tyr Phe Asn Gln Leu Gln Ala Met 290 295 300 His Thr Leu Gln Asp Thr Ser Trp His Asn Thr Thr Phe Phe Gly 305 310 315 Val Phe Gln Ala Gln Trp Gly Asp Met Tyr Leu Ser Ala Ile Cys 320 325 330 Glu Tyr Gln Leu Glu Glu Ile Gln Arg Val Phe Glu Gly Pro Tyr 335 340 345 Lys Glu Tyr His Glu Glu Ala Gln Lys Trp Asp Arg Tyr Thr Asp 350 355 360 Pro Val Pro Ser Pro Arg Pro Gly Ser Cys Ile Asn Asn Trp His 365 370 375 Arg Arg His Gly Tyr Thr Ser Ser Leu Glu Leu Pro Asp Asn Ile 380 385 390 Leu Asn Phe Val Lys Lys His Pro Leu Met Glu Glu Gln Val Gly 395 400 405 Pro Arg Trp Ser Arg Pro Leu Leu Val Lys Lys Gly Thr Asn Phe 410 415 420 Thr His Leu Val Ala Asp Arg Val Thr Gly Leu Asp Gly Ala Thr 425 430 435 Tyr Thr Val Leu Phe Ile Gly Thr Gly Asp Gly Trp Leu Leu Lys 440 445 450 Ala Val Ser Leu Gly Pro Trp Val His Leu Ile Glu Glu Leu Gln 455 460 465 Leu Phe Asp Gln Glu Pro Met Arg Ser Leu Val Leu Ser Gln Ser 470 475 480 Lys Lys Leu Leu Phe Ala Gly Ser Arg Ser Gln Leu Val Gln Leu 485 490 495 Pro Val Ala Asp Cys Met Lys Tyr Arg Ser Cys Ala Asp Cys Val 500 505 510 Leu Ala Arg Asp Pro Tyr Cys Ala Trp Ser Val Asn Thr Ser Arg 515 520 525 Cys Val Ala Val Gly Gly His Ser Gly Ser Leu Leu Ile Gln His 530 535 540 Val Met Thr Ser Asp Thr Ser Gly Ile Cys Asn Leu Arg Gly Ser 545 550 555 Lys Lys Val Arg Pro Thr Pro Lys Asn Ile Thr Val Val Ala Gly 560 565 570 Thr Asp Leu Val Leu Pro Cys His Leu Ser Ser Asn Leu Ala His 575 580 585 Ala Arg Trp Thr Phe Gly Gly Arg Asp Leu Pro Ala Glu Gln Pro 590 595 600 Gly Ser Phe Leu Tyr Asp Ala Arg Leu Gln Ala Leu Val Val Met 605 610 615 Ala Ala Gln Pro Arg His Ala Gly Ala Tyr His Cys Phe Ser Glu 620 625 630 Glu Gln Gly Ala Arg Leu Ala Ala Glu Gly Tyr Leu Val Ala Val 635 640 645 Val Ala Gly Pro Ser Val Thr Leu Glu Ala Arg Ala Pro Leu Glu 650 655 660 Asn Leu Gly Leu Val Trp Leu Ala Val Val Ala Leu Gly Ala Val 665 670 675 Cys Leu Val Leu Leu Leu Leu Val Leu Ser Leu Arg Arg Arg Leu 680 685 690 Arg Glu Glu Leu Glu Lys Gly Ala Lys Ala Thr Glu Arg Thr Leu 695 700 705 Val Tyr Pro Leu Glu Leu Pro Lys Glu Pro Thr Ser Pro Pro Phe 710 715 720 Arg Pro Cys Pro Glu Pro Asp Glu Lys Leu Trp Asp Pro Val Gly 725 730 735 Tyr Tyr Tyr Ser Asp Gly Ser Leu Lys Ile Val Pro Gly His Ala 740 745 750 Arg Cys Gln Pro Gly Gly Gly Pro Pro Ser Pro Pro Pro Gly Ile 755 760 765 Pro Gly Gln Pro Leu Pro Ser Pro Thr Arg Leu His Leu Gly Gly 770 775 780 Gly Arg Asn Ser Asn Ala Asn Gly Tyr Val Arg Leu Gln Leu Gly 785 790 795 Gly Glu Asp Arg Gly Gly Leu Gly His Pro Leu Pro Glu Leu Ala 800 805 810 Asp Glu Leu Arg Arg Lys Leu Gln Gln Arg Gln Pro Leu Pro Asp 815 820 825 Ser Asn Pro Glu Glu Ser Ser Val 830 5 410 PRT Homo sapiens misc_feature Incyte ID No 1689337CD1 5 Met Ser Asn Ile Ser Leu Leu Ala Leu Phe Ser Ser Gln Val Ser 1 5 10 15 Ala Ser Leu Lys Ala Leu Ser His Phe Phe Ser Leu Cys Phe Arg 20 25 30 Leu Ala Arg Glu Gln Ala Arg Val Cys Glu Leu Gln Ser Gly Asn 35 40 45 Gln Gln Leu Glu Glu Gln Arg Val Glu Leu Val Glu Arg Leu Gln 50 55 60 Ala Met Leu Gln Ala His Trp Asp Glu Ala Asn Gln Leu Leu Ser 65 70 75 Thr Thr Leu Pro Pro Pro Asn Pro Pro Ala Pro Pro Ala Gly Pro 80 85 90 Ser Ser Pro Gly Pro Gln Glu Pro Glu Lys Glu Glu Arg Arg Val 95 100 105 Trp Thr Met Pro Pro Met Ala Val Ala Leu Lys Pro Val Leu Gln 110 115 120 Gln Ser Arg Glu Ala Arg Asp Glu Leu Pro Gly Ala Pro Pro Val 125 130 135 Leu Cys Ser Ser Ser Ser Asp Leu Ser Leu Leu Leu Gly Pro Ser 140 145 150 Phe Gln Ser Gln His Ser Phe Gln Pro Leu Glu Pro Lys Pro Asp 155 160 165 Leu Thr Ser Ser Thr Ala Gly Ala Phe Ser Ala Leu Gly Ala Phe 170 175 180 His Pro Asp His Arg Ala Glu Arg Pro Phe Pro Glu Glu Asp Pro 185 190 195 Gly Pro Asp Gly Glu Gly Leu Leu Lys Gln Gly Leu Pro Pro Ala 200 205 210 Gln Leu Glu Gly Leu Lys Asn Phe Leu His Gln Leu Leu Glu Thr 215 220 225 Val Pro Gln Asn Asn Glu Asn Pro Ser Val Asp Leu Leu Pro Pro 230 235 240 Lys Ser Gly Pro Leu Thr Val Pro Ser Trp Glu Glu Ala Pro Gln 245 250 255 Val Pro Arg Ile Pro Pro Pro Val His Lys Thr Lys Val Pro Leu 260 265 270 Ala Met Ala Ser Ser Leu Phe Arg Val Pro Glu Pro Pro Ser Ser 275 280 285 His Ser Gln Gly Ser Gly Pro Ser Ser Gly Ser Pro Glu Arg Gly 290 295 300 Gly Asp Gly Leu Thr Phe Pro Arg Gln Leu Met Glu Val Ser Gln 305 310 315 Leu Leu Arg Leu Tyr Gln Ala Arg Gly Trp Gly Ala Leu Pro Ala 320 325 330 Glu Asp Leu Leu Leu Tyr Leu Lys Arg Leu Glu His Ser Gly Arg 335 340 345 Thr Asp Gly Arg Gly Asp Asn Val Pro Arg Arg Asn Thr Asp Ser 350 355 360 Arg Leu Gly Glu Ile Pro Arg Lys Glu Ile Pro Ser Gln Ala Val 365 370 375 Pro Arg Arg Leu Ala Thr Ala Pro Lys Thr Glu Lys Pro Pro Ala 380 385 390 Arg Lys Lys Ser Gly His Pro Ala Pro Ser Ser Met Arg Ser Arg 395 400 405 Gly Gly Val Trp Arg 410 6 360 PRT Homo sapiens misc_feature Incyte ID No 1746392CD1 6 Met Asp Thr Pro Leu Arg Arg Ser Arg Arg Leu Gly Gly Leu Arg 1 5 10 15 Pro Glu Ser Pro Glu Ser Leu Thr Ser Val Ser Arg Thr Arg Arg 20 25 30 Ala Leu Val Glu Phe Glu Ser Asn Pro Glu Glu Thr Arg Glu Pro 35 40 45 Gly Ser Pro Pro Ser Val Gln Arg Ala Gly Leu Gly Ser Pro Glu 50 55 60 Arg Pro Pro Lys Thr Ser Pro Gly Ser Pro Arg Leu Gln Gln Gly 65 70 75 Ala Gly Leu Glu Ser Pro Gln Gly Gln Pro Glu Pro Gly Ala Ala 80 85 90 Ser Pro Gln Arg Gln Gln Asp Leu His Leu Glu Ser Pro Gln Arg 95 100 105 Gln Pro Glu Tyr Ser Pro Glu Ser Pro Arg Cys Gln Pro Lys Pro 110 115 120 Ser Glu Glu Ala Pro Lys Cys Ser Gln Asp Gln Gly Val Leu Ala 125 130 135 Ser Glu Leu Ala Gln Asn Lys Glu Glu Leu Thr Pro Gly Ala Pro 140 145 150 Gln His Gln Leu Pro Pro Val Pro Gly Ser Pro Glu Pro Tyr Pro 155 160 165 Gly Gln Gln Ala Pro Gly Pro Glu Pro Ser Gln Pro Leu Leu Glu 170 175 180 Leu Thr Pro Arg Ala Pro Gly Ser Pro Arg Gly Gln His Glu Pro 185 190 195 Ser Lys Pro Pro Pro Ala Gly Glu Thr Val Thr Gly Gly Phe Gly 200 205 210 Ala Lys Lys Arg Lys Gly Ser Ser Ser Gln Ala Pro Ala Ser Lys 215 220 225 Lys Leu Asn Lys Glu Glu Leu Pro Val Ile Pro Lys Gly Lys Pro 230 235 240 Lys Ser Gly Arg Val Trp Lys Asp Arg Ser Lys Lys Arg Phe Ser 245 250 255 Gln Met Leu Gln Asp Lys Pro Leu Arg Thr Ser Trp Gln Arg Lys 260 265 270 Met Lys Glu Arg Gln Glu Arg Lys Leu Ala Lys Asp Phe Ala Arg 275 280 285 His Leu Glu Glu Glu Lys Glu Arg Arg Arg Gln Glu Lys Lys Gln 290 295 300 Arg Arg Ala Glu Asn Leu Lys Arg Arg Leu Glu Asn Glu Arg Lys 305 310 315 Ala Glu Val Val Gln Val Ile Arg Asn Pro Ala Lys Leu Lys Arg 320 325 330 Ala Lys Lys Lys Gln Leu Arg Ser Ile Glu Lys Arg Asp Thr Leu 335 340 345 Ala Leu Leu Gln Lys Gln Pro Pro Gln Gln Pro Ala Ala Lys Ile 350 355 360 7 377 PRT Homo sapiens misc_feature Incyte ID No 1825182CD1 7 Met Lys Thr Leu Pro Leu Phe Val Cys Ile Cys Ala Leu Ser Ala 1 5 10 15 Cys Phe Ser Phe Ser Glu Gly Arg Glu Arg Asp His Glu Leu Arg 20 25 30 His Arg Arg His His His Gln Ser Pro Lys Ser His Phe Glu Leu 35 40 45 Pro His Tyr Pro Gly Leu Leu Ala His Gln Lys Pro Phe Ile Arg 50 55 60 Lys Ser Tyr Lys Cys Leu His Lys Arg Cys Arg Pro Lys Leu Pro 65 70 75 Pro Ser Pro Asn Lys Pro Pro Lys Phe Pro Asn Pro His Gln Pro 80 85 90 Pro Lys His Pro Asp Lys Asn Ser Ser Val Val Asn Pro Thr Leu 95 100 105 Val Ala Thr Thr Gln Ile Pro Ser Val Thr Phe Pro Ser Ala Ser 110 115 120 Thr Lys Ile Thr Thr Leu Pro Asn Val Thr Phe Leu Pro Gln Asn 125 130 135 Ala Thr Thr Ile Ser Ser Arg Glu Asn Val Asn Thr Ser Ser Ser 140 145 150 Val Ala Thr Leu Ala Pro Val Asn Ser Pro Ala Pro Gln Asp Thr 155 160 165 Thr Ala Ala Pro Pro Thr Pro Ser Ala Thr Thr Pro Ala Pro Pro 170 175 180 Ser Ser Ser Ala Pro Pro Glu Thr Thr Ala Ala Pro Pro Thr Pro 185 190 195 Ser Ala Thr Thr Gln Ala Pro Pro Ser Ser Ser Ala Pro Pro Glu 200 205 210 Thr Thr Ala Ala Pro Pro Thr Pro Pro Ala Thr Thr Pro Ala Pro 215 220 225 Pro Ser Ser Ser Ala Pro Pro Glu Thr Thr Ala Ala Pro Pro Thr 230 235 240 Pro Ser Ala Thr Thr Pro Ala Pro Leu Ser Ser Ser Ala Pro Pro 245 250 255 Glu Thr Thr Ala Val Pro Pro Thr Pro Ser Ala Thr Thr Leu Asp 260 265 270 Pro Ser Ser Ala Ser Ala Pro Pro Glu Thr Thr Ala Ala Pro Pro 275 280 285 Thr Pro Ser Ala Thr Thr Pro Ala Pro Pro Ser Ser Pro Ala Pro 290 295 300 Gln Glu Thr Thr Ala Ala Pro Ile Thr Thr Pro Asn Ser Ser Pro 305 310 315 Thr Thr Leu Ala Pro Asp Thr Ser Glu Thr Ser Ala Ala Pro Thr 320 325 330 His Gln Thr Thr Thr Ser Val Thr Thr Gln Thr Thr Thr Thr Lys 335 340 345 Gln Pro Thr Ser Ala Pro Gly Gln Asn Lys Ile Ser Arg Phe Leu 350 355 360 Leu Tyr Met Lys Asn Leu Leu Asn Arg Ile Ile Asp Asp Met Val 365 370 375 Glu Gln 8 182 PRT Homo sapiens misc_feature Incyte ID No 2155541CD1 8 Met Leu Val Leu Val Leu Gly Asp Leu His Ile Pro His Arg Cys 1 5 10 15 Asn Ser Leu Pro Ala Lys Phe Lys Lys Leu Leu Val Pro Gly Lys 20 25 30 Ile Gln His Ile Leu Cys Thr Gly Asn Leu Cys Thr Lys Glu Ser 35 40 45 Tyr Asp Tyr Leu Lys Thr Leu Ala Gly Asp Val His Ile Val Arg 50 55 60 Gly Asp Phe Asp Glu Asn Leu Asn Tyr Pro Glu Gln Lys Val Val 65 70 75 Thr Val Gly Gln Phe Lys Ile Gly Leu Ile His Gly His Gln Val 80 85 90 Ile Pro Trp Gly Asp Met Ala Ser Leu Ala Leu Leu Gln Arg Gln 95 100 105 Phe Asp Val Asp Ile Leu Ile Ser Gly His Thr His Lys Phe Glu 110 115 120 Ala Phe Glu His Glu Asn Lys Phe Tyr Ile Asn Pro Gly Ser Ala 125 130 135 Thr Gly Ala Tyr Asn Ala Leu Glu Thr Asn Ile Ile Pro Ser Phe 140 145 150 Val Leu Met Asp Ile Gln Ala Ser Thr Val Val Thr Tyr Val Tyr 155 160 165 Gln Leu Ile Gly Asp Asp Val Lys Val Glu Arg Ile Glu Tyr Lys 170 175 180 Lys Pro 9 513 PRT Homo sapiens misc_feature Incyte ID No 2215706CD1 9 Met Asn Met Asn Phe Gly Asp Trp His Leu Phe Arg Ser Thr Val 1 5 10 15 Leu Glu Met Arg Asn Ala Glu Ser His Val Val Pro Glu Asp Pro 20 25 30 Arg Phe Leu Ser Glu Ser Ser Ser Gly Pro Ala Pro His Gly Glu 35 40 45 Pro Ala Arg Arg Ala Ser His Asn Glu Leu Pro His Thr Glu Leu 50 55 60 Ser Ser Gln Thr Pro Tyr Thr Leu Asn Phe Ser Phe Glu Glu Leu 65 70 75 Asn Thr Leu Gly Leu Asp Glu Gly Ala Pro Arg His Ser Asn Leu 80 85 90 Ser Trp Gln Ser Gln Thr Arg Arg Thr Pro Ser Leu Ser Ser Leu 95 100 105 Asn Ser Gln Asp Ser Ser Ile Glu Ile Ser Lys Leu Thr Asp Lys 110 115 120 Val Gln Ala Glu Tyr Arg Asp Ala Tyr Arg Glu Tyr Ile Ala Gln 125 130 135 Met Ser Gln Leu Glu Gly Gly Pro Gly Ser Thr Thr Ile Ser Gly 140 145 150 Arg Ser Ser Pro His Ser Thr Tyr Tyr Met Gly Gln Ser Ser Ser 155 160 165 Gly Gly Ser Ile His Ser Asn Leu Glu Gln Glu Lys Gly Lys Asp 170 175 180 Ser Glu Pro Lys Pro Asp Asp Gly Arg Lys Ser Phe Leu Met Lys 185 190 195 Arg Gly Asp Val Ile Asp Tyr Ser Ser Ser Gly Val Ser Thr Asn 200 205 210 Asp Ala Ser Pro Leu Asp Pro Ile Thr Glu Glu Asp Glu Lys Ser 215 220 225 Asp Gln Ser Gly Ser Lys Leu Leu Pro Gly Lys Lys Ser Ser Glu 230 235 240 Arg Ser Ser Leu Phe Gln Thr Asp Leu Lys Leu Lys Gly Ser Gly 245 250 255 Leu Arg Tyr Gln Lys Leu Pro Ser Asp Glu Asp Glu Ser Gly Thr 260 265 270 Glu Glu Ser Asp Asn Thr Pro Leu Leu Lys Asp Asp Lys Asp Arg 275 280 285 Lys Ala Glu Gly Lys Val Glu Arg Val Pro Lys Ser Pro Glu His 290 295 300 Ser Ala Glu Pro Ile Arg Thr Phe Ile Lys Ala Lys Glu Tyr Leu 305 310 315 Ser Asp Ala Leu Leu Asp Lys Lys Asp Ser Ser Asp Ser Gly Val 320 325 330 Arg Ser Ser Glu Ser Ser Pro Asn His Ser Leu His Asn Glu Val 335 340 345 Ala Asp Asp Ser His Leu Glu Lys Ala Asn Leu Ile Glu Leu Glu 350 355 360 Asp Asp Ser His Ser Gly Lys Arg Gly Ile Pro His Ser Leu Ser 365 370 375 Gly Leu Gln Asp Pro Ile Ile Ala Arg Met Ser Ile Cys Ser Glu 380 385 390 Asp Lys Lys Ser Pro Ser Glu Cys Ser Leu Ile Ala Ser Ser Pro 395 400 405 Glu Glu Asn Trp Pro Ala Cys Gln Lys Ala Tyr Asn Leu Asn Arg 410 415 420 Thr Pro Ser Thr Val Thr Leu Asn Asn Asn Ser Ala Pro Ala Asn 425 430 435 Arg Ala Asn Gln Asn Phe Asp Glu Met Glu Gly Ile Arg Glu Thr 440 445 450 Ser Gln Val Ile Leu Arg Pro Ser Ser Ser Pro Asn Pro Thr Thr 455 460 465 Ile Gln Asn Glu Asn Leu Lys Ser Met Thr His Lys Arg Ser Gln 470 475 480 Arg Ser Ser Tyr Thr Arg Leu Ser Lys Asp Pro Pro Glu Leu His 485 490 495 Ala Ala Ala Ser Ser Glu Ser Thr Gly Phe Gly Glu Glu Arg Glu 500 505 510 Ser Ile Leu 10 361 PRT Homo sapiens misc_feature Incyte ID No 2347692CD1 10 Met Tyr Gly Lys Gly Lys Ser Asn Ser Ser Ala Val Pro Ser Asp 1 5 10 15 Ser Gln Ala Arg Glu Lys Leu Ala Leu Tyr Val Tyr Glu Tyr Leu 20 25 30 Leu His Val Gly Ala Gln Lys Ser Ala Gln Thr Phe Leu Ser Glu 35 40 45 Ile Arg Trp Glu Lys Asn Ile Thr Leu Gly Glu Pro Pro Gly Phe 50 55 60 Leu His Ser Trp Trp Cys Val Phe Trp Asp Leu Tyr Cys Ala Ala 65 70 75 Pro Glu Arg Arg Glu Thr Cys Glu His Ser Ser Glu Ala Lys Ala 80 85 90 Phe His Asp Tyr Ser Ala Ala Ala Ala Pro Ser Pro Val Leu Gly 95 100 105 Asn Ile Pro Pro Gly Asp Gly Met Pro Val Gly Pro Val Pro Pro 110 115 120 Gly Phe Phe Gln Pro Phe Met Ser Pro Arg Tyr Pro Gly Gly Pro 125 130 135 Arg Pro Pro Leu Arg Ile Pro Asn Gln Ala Leu Gly Gly Val Pro 140 145 150 Gly Ser Gln Pro Leu Leu Pro Ser Gly Met Asp Pro Thr Arg Gln 155 160 165 Gln Gly His Pro Asn Met Gly Gly Pro Met Gln Arg Met Thr Pro 170 175 180 Pro Arg Gly Met Val Pro Leu Gly Pro Gln Asn Tyr Gly Gly Ala 185 190 195 Met Arg Pro Pro Leu Asn Ala Leu Gly Gly Pro Gly Met Pro Gly 200 205 210 Met Asn Met Gly Pro Gly Gly Gly Arg Pro Trp Pro Asn Pro Thr 215 220 225 Asn Ala Asn Ser Ile Pro Tyr Ser Ser Ala Ser Pro Gly Asn Tyr 230 235 240 Val Gly Pro Pro Gly Gly Gly Gly Pro Pro Gly Thr Pro Ile Met 245 250 255 Pro Ser Pro Ala Asp Ser Thr Asn Ser Gly Asp Asn Met Tyr Thr 260 265 270 Leu Met Asn Ala Val Pro Pro Gly Pro Asn Arg Pro Asn Phe Pro 275 280 285 Met Gly Pro Gly Ser Asp Gly Pro Met Gly Gly Leu Gly Gly Met 290 295 300 Glu Ser His His Met Asn Gly Ser Leu Gly Ser Gly Asp Met Asp 305 310 315 Ser Ile Ser Lys Asn Ser Pro Asn Asn Met Ser Leu Ser Asn Gln 320 325 330 Pro Gly Thr Pro Arg Asp Asp Gly Glu Met Gly Gly Asn Phe Leu 335 340 345 Asn Pro Phe Gln Ser Glu Ser Tyr Ser Pro Ser Met Thr Met Ser 350 355 360 Val 11 327 PRT Homo sapiens misc_feature Incyte ID No 2579048CD1 11 Met Ala Leu Val His Lys Leu Leu His Gly Thr Tyr Phe Leu Arg 1 5 10 15 Lys Phe Ser Lys Pro Thr Ser Ala Leu Tyr Pro Phe Leu Gly Ile 20 25 30 Leu Phe Ala Glu Tyr Ser Ser Ser Leu Gln Lys Pro Val Ala Ser 35 40 45 Pro Gly Lys Ala Ser Ser Gln Arg Lys Thr Glu Gly Asp Leu Gln 50 55 60 Gly Asp His Gln Lys Glu Val Ala Leu Asp Ile Thr Ser Ser Glu 65 70 75 Glu Lys Pro Asp Val Ser Phe Asp Lys Ala Ile Arg Asp Glu Ala 80 85 90 Ile Tyr His Phe Arg Leu Leu Lys Asp Glu Ile Val Asp His Trp 95 100 105 Arg Gly Pro Glu Gly His Pro Leu His Glu Val Leu Leu Glu Gln 110 115 120 Ala Lys Val Val Trp Gln Phe Arg Gly Lys Glu Asp Leu Asp Lys 125 130 135 Trp Thr Val Thr Ser Asp Lys Thr Ile Gly Gly Arg Ser Glu Val 140 145 150 Phe Leu Lys Met Gly Lys Asn Asn Gln Ser Ala Leu Leu Tyr Gly 155 160 165 Thr Leu Ser Ser Glu Ala Pro Gln Asp Gly Glu Ser Thr Arg Ser 170 175 180 Gly Tyr Cys Ala Met Ile Ser Arg Ile Pro Arg Gly Ala Phe Glu 185 190 195 Arg Lys Met Ser Tyr Asp Trp Ser Gln Phe Asn Thr Leu Tyr Leu 200 205 210 Arg Val Arg Gly Asp Gly Arg Pro Trp Met Val Asn Ile Lys Glu 215 220 225 Asp Thr Asp Phe Phe Gln Arg Thr Asn Gln Met Tyr Ser Tyr Phe 230 235 240 Met Phe Thr Arg Gly Gly Pro Tyr Trp Gln Glu Val Lys Ile Pro 245 250 255 Phe Ser Lys Phe Phe Phe Ser Asn Arg Gly Arg Ile Arg Asp Val 260 265 270 Gln His Glu Leu Pro Leu Asp Lys Ile Ser Ser Ile Gly Phe Thr 275 280 285 Leu Ala Asp Lys Val Asp Gly Pro Phe Phe Leu Glu Ile Asp Phe 290 295 300 Ile Gly Val Phe Thr Asp Pro Ala His Thr Glu Glu Phe Ala Tyr 305 310 315 Glu Asn Ser Pro Glu Leu Asn Pro Arg Leu Phe Lys 320 325 12 1110 PRT Homo sapiens misc_feature Incyte ID No 2604493CD1 12 Met Pro Ala Pro Glu Gln Ala Ser Leu Val Glu Glu Gly Gln Pro 1 5 10 15 Gln Thr Arg Gln Glu Ala Ala Ser Thr Gly Pro Gly Met Glu Pro 20 25 30 Glu Thr Thr Ala Thr Thr Ile Leu Ala Ser Val Lys Glu Gln Glu 35 40 45 Leu Gln Phe Gln Arg Leu Thr Arg Glu Leu Glu Val Glu Arg Gln 50 55 60 Ile Val Ala Ser Gln Leu Glu Arg Cys Arg Leu Gly Ala Glu Ser 65 70 75 Pro Ser Ile Ala Ser Thr Ser Ser Thr Glu Lys Ser Phe Pro Trp 80 85 90 Arg Ser Thr Asp Val Pro Asn Thr Gly Val Ser Lys Pro Arg Val 95 100 105 Ser Asp Ala Val Gln Pro Asn Asn Tyr Leu Ile Arg Thr Glu Pro 110 115 120 Glu Gln Gly Thr Leu Tyr Ser Pro Glu Gln Thr Ser Leu His Glu 125 130 135 Ser Glu Gly Ser Leu Gly Asn Ser Arg Ser Ser Thr Gln Met Asn 140 145 150 Ser Tyr Ser Asp Ser Gly Tyr Gln Glu Ala Gly Ser Phe His Asn 155 160 165 Ser Gln Asn Val Ser Lys Ala Asp Asn Arg Gln Gln His Ser Phe 170 175 180 Ile Gly Ser Thr Asn Asn His Val Val Arg Asn Ser Arg Ala Glu 185 190 195 Gly Gln Thr Leu Val Gln Pro Ser Val Ala Asn Arg Ala Met Arg 200 205 210 Arg Val Ser Ser Val Pro Ser Arg Ala Gln Ser Pro Ser Tyr Val 215 220 225 Ile Ser Thr Gly Val Ser Pro Ser Arg Gly Ser Leu Arg Thr Ser 230 235 240 Leu Gly Ser Gly Phe Gly Ser Pro Ser Val Thr Asp Pro Arg Pro 245 250 255 Leu Asn Pro Ser Ala Tyr Ser Ser Thr Thr Leu Pro Ala Ala Arg 260 265 270 Ala Ala Ser Pro Tyr Ser Gln Arg Pro Ala Ser Pro Thr Ala Ile 275 280 285 Arg Arg Ile Gly Ser Val Thr Ser Arg Gln Thr Ser Asn Pro Asn 290 295 300 Gly Pro Thr Pro Gln Tyr Gln Thr Thr Ala Arg Val Gly Ser Pro 305 310 315 Leu Thr Leu Thr Asp Ala Gln Thr Arg Val Ala Ser Pro Ser Gln 320 325 330 Gly Gln Val Gly Ser Ser Ser Pro Lys Arg Ser Gly Met Thr Ala 335 340 345 Val Pro Gln His Leu Gly Pro Ser Leu Gln Arg Thr Val His Asp 350 355 360 Met Glu Gln Phe Gly Gln Gln Gln Tyr Asp Ile Tyr Glu Arg Met 365 370 375 Val Pro Pro Arg Pro Asp Ser Leu Thr Gly Leu Arg Ser Ser Tyr 380 385 390 Ala Ser Gln His Ser Gln Leu Gly Gln Asp Leu Arg Ser Ala Val 395 400 405 Ser Pro Asp Leu His Ile Thr Pro Ile Tyr Glu Gly Arg Thr Tyr 410 415 420 Tyr Ser Pro Val Tyr Arg Ser Pro Asn His Gly Thr Val Glu Leu 425 430 435 Gln Gly Ser Gln Thr Ala Leu Tyr Arg Thr Gly Ser Gly Ile Gly 440 445 450 Asn Leu Gln Arg Thr Ser Ser Gln Arg Ser Thr Leu Thr Tyr Gln 455 460 465 Arg Asn Asn Tyr Ala Leu Asn Thr Thr Ala Thr Tyr Ala Glu Pro 470 475 480 Tyr Arg Pro Ile Gln Tyr Arg Val Gln Glu Cys Asn Tyr Asn Arg 485 490 495 Leu Gln His Ala Val Pro Ala Asp Asp Gly Thr Thr Arg Ser Pro 500 505 510 Ser Ile Asp Ser Ile Gln Lys Asp Pro Arg Glu Phe Ala Trp Arg 515 520 525 Asp Pro Glu Leu Pro Glu Val Ile His Met Leu Gln His Gln Phe 530 535 540 Pro Ser Val Gln Ala Asn Ala Ala Ala Tyr Leu Gln His Leu Cys 545 550 555 Phe Gly Asp Asn Lys Val Lys Met Glu Val Cys Arg Leu Gly Gly 560 565 570 Ile Lys His Leu Val Asp Leu Leu Asp His Arg Val Leu Glu Val 575 580 585 Gln Lys Asn Ala Cys Gly Ala Leu Arg Asn Leu Val Phe Gly Lys 590 595 600 Ser Thr Asp Glu Asn Lys Ile Ala Met Lys Asn Val Gly Gly Ile 605 610 615 Pro Ala Leu Leu Arg Leu Leu Arg Lys Ser Ile Asp Ala Glu Val 620 625 630 Arg Glu Leu Val Thr Gly Val Leu Trp Asn Leu Ser Ser Cys Asp 635 640 645 Ala Val Lys Met Thr Ile Ile Arg Asp Ala Leu Ser Thr Leu Thr 650 655 660 Asn Thr Val Ile Val Pro His Ser Gly Trp Asn Asn Ser Ser Phe 665 670 675 Asp Asp Asp His Lys Ile Lys Phe Gln Thr Ser Leu Val Leu Arg 680 685 690 Asn Thr Thr Gly Cys Leu Arg Asn Leu Ser Ser Ala Gly Glu Glu 695 700 705 Ala Arg Lys Gln Met Arg Ser Cys Glu Gly Leu Val Asp Ser Leu 710 715 720 Leu Tyr Val Ile His Thr Cys Val Asn Thr Ser Asp Tyr Asp Ser 725 730 735 Lys Thr Val Glu Asn Cys Val Cys Thr Leu Arg Asn Leu Ser Tyr 740 745 750 Arg Leu Glu Leu Glu Val Pro Gln Ala Arg Leu Leu Gly Leu Asn 755 760 765 Glu Leu Asp Asp Leu Leu Gly Lys Glu Ser Pro Ser Lys Asp Ser 770 775 780 Glu Pro Ser Cys Trp Gly Lys Lys Lys Lys Lys Lys Lys Arg Thr 785 790 795 Pro Gln Glu Asp Gln Trp Asp Gly Val Gly Pro Ile Pro Gly Leu 800 805 810 Ser Lys Ser Pro Lys Gly Val Glu Met Leu Trp His Pro Ser Val 815 820 825 Val Lys Pro Tyr Leu Thr Leu Leu Ala Glu Ser Ser Asn Pro Ala 830 835 840 Thr Leu Glu Gly Ser Ala Gly Ser Leu Gln Asn Leu Ser Ala Gly 845 850 855 Asn Trp Lys Phe Ala Ala Tyr Ile Arg Ala Ala Val Arg Lys Glu 860 865 870 Lys Gly Leu Pro Ile Leu Val Glu Leu Leu Arg Met Asp Asn Asp 875 880 885 Arg Val Val Ser Ser Val Ala Thr Ala Leu Arg Asn Met Ala Leu 890 895 900 Asp Val Arg Asn Lys Glu Leu Ile Gly Lys Tyr Ala Met Arg Asp 905 910 915 Leu Val Asn Arg Leu Pro Gly Gly Asn Gly Pro Ser Val Leu Ser 920 925 930 Asp Glu Thr Met Ala Ala Ile Cys Cys Ala Leu His Glu Val Thr 935 940 945 Ser Lys Asn Met Glu Asn Ala Lys Ala Leu Ala Asp Ser Gly Gly 950 955 960 Ile Glu Lys Leu Val Asn Ile Thr Lys Gly Arg Gly Asp Arg Ser 965 970 975 Ser Leu Lys Val Val Lys Ala Ala Ala Gln Val Leu Asn Thr Leu 980 985 990 Trp Gln Tyr Arg Asp Leu Arg Ser Ile Tyr Lys Lys Asp Gly Trp 995 1000 1005 Asn Gln Asn His Phe Ile Thr Pro Val Ser Thr Leu Glu Arg Asp 1010 1015 1020 Arg Phe Lys Ser His Pro Ser Leu Ser Thr Thr Asn Gln Gln Met 1025 1030 1035 Ser Pro Ile Ile Gln Ser Val Gly Ser Thr Ser Ser Ser Pro Ala 1040 1045 1050 Leu Leu Gly Ile Arg Asp Pro Arg Ser Glu Tyr Asp Arg Thr Gln 1055 1060 1065 Pro Pro Met Gln Tyr Tyr Asn Ser Gln Gly Asp Ala Thr His Lys 1070 1075 1080 Gly Leu Tyr Pro Gly Lys Thr Pro Val Gly Cys Val Ile Gln Ser 1085 1090 1095 Leu Glu Lys Pro His Phe Gln Ala Leu Gly Gln Trp Pro Gly Lys 1100 1105 1110 13 386 PRT Homo sapiens misc_feature Incyte ID No 2787182CD1 13 Met Asp Arg Phe Val Trp Thr Ser Gly Leu Leu Glu Ile Asn Glu 1 5 10 15 Thr Leu Val Ile Gln Gln Arg Gly Val Arg Ile Tyr Asp Gly Glu 20 25 30 Glu Lys Ile Lys Phe Asp Ala Gly Thr Leu Leu Leu Ser Thr His 35 40 45 Arg Leu Ile Trp Arg Asp Gln Lys Asn His Glu Cys Cys Met Ala 50 55 60 Ile Leu Leu Ser Gln Ile Val Phe Ile Glu Glu Gln Ala Ala Gly 65 70 75 Ile Gly Lys Ser Ala Lys Ile Val Val His Leu His Pro Ala Pro 80 85 90 Pro Asn Lys Glu Pro Gly Pro Phe Gln Ser Ser Lys Asn Ser Tyr 95 100 105 Ile Lys Leu Ser Phe Lys Glu His Gly Gln Ile Glu Phe Tyr Arg 110 115 120 Arg Leu Ser Glu Glu Met Thr Gln Arg Arg Trp Glu Asn Met Pro 125 130 135 Val Ser Gln Ser Leu Gln Thr Asn Arg Gly Pro Gln Pro Gly Arg 140 145 150 Ile Arg Ala Val Gly Ile Val Gly Ile Glu Arg Lys Leu Glu Glu 155 160 165 Lys Lys Lys Glu Thr Asp Lys Asn Ile Ser Glu Ala Phe Glu Asp 170 175 180 Leu Ser Lys Leu Met Ile Lys Ala Lys Glu Met Val Glu Leu Ser 185 190 195 Lys Ser Ile Ala Asn Lys Ile Lys Asp Lys Gln Gly Asp Ile Thr 200 205 210 Glu Asp Glu Thr Ile Arg Phe Lys Ser Tyr Leu Leu Ser Met Gly 215 220 225 Ile Ala Asn Pro Val Thr Arg Glu Thr Tyr Gly Ser Gly Thr Gln 230 235 240 Tyr His Met Gln Leu Ala Lys Gln Leu Ala Gly Ile Leu Gln Val 245 250 255 Pro Leu Glu Glu Arg Gly Gly Ile Met Ser Leu Thr Glu Val Tyr 260 265 270 Cys Leu Val Asn Arg Ala Arg Gly Met Glu Leu Leu Ser Pro Glu 275 280 285 Asp Leu Val Asn Ala Cys Lys Met Leu Glu Ala Leu Lys Leu Pro 290 295 300 Leu Arg Leu Arg Val Phe Asp Ser Gly Val Met Val Ile Glu Leu 305 310 315 Gln Ser His Lys Glu Glu Glu Met Val Ala Ser Ala Leu Glu Thr 320 325 330 Val Ser Glu Lys Gly Ser Leu Thr Ser Glu Glu Phe Ala Lys Leu 335 340 345 Val Gly Met Ser Val Leu Leu Ala Lys Glu Arg Leu Leu Leu Ala 350 355 360 Glu Lys Met Gly His Leu Cys Arg Asp Asp Ser Val Glu Gly Leu 365 370 375 Arg Phe Tyr Pro Asn Leu Phe Met Thr Gln Ser 380 385 14 181 PRT Homo sapiens misc_feature Incyte ID No 3096668CD1 14 Met Glu Val Val Glu Ala Ala Ala Ala Gln Leu Glu Thr Leu Lys 1 5 10 15 Phe Asn Gly Thr Asp Phe Gly Val Gly Glu Gly Pro Ala Ala Pro 20 25 30 Ser Pro Gly Ser Ala Pro Val Pro Gly Thr Gln Pro Pro Leu Gln 35 40 45 Ser Phe Glu Gly Ser Pro Asp Ala Gly Gln Thr Val Glu Val Lys 50 55 60 Pro Ala Gly Glu Gln Pro Leu Gln Pro Val Leu Asn Ala Val Ala 65 70 75 Ala Gly Thr Pro Ala Pro Gln Pro Gln Pro Pro Ala Glu Ser Pro 80 85 90 Ala Cys Gly Asp Cys Val Thr Ser Pro Gly Ala Ala Glu Pro Ala 95 100 105 Arg Ala Pro Asp Ser Leu Glu Thr Ser Asp Ser Asp Ser Asp Ser 110 115 120 Asp Ser Glu Thr Asp Ser Asp Ser Ser Ser Ser Ser Ser Ser Ser 125 130 135 Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Cys Ile Ser Leu Pro 140 145 150 Pro Val Leu Ser Asp Gly Asp Asp Asp Leu Gln Val Glu Lys Glu 155 160 165 Asn Lys Asn Phe Pro Leu Lys Thr Lys Asp Glu Leu Leu Leu Asn 170 175 180 Leu 15 374 PRT Homo sapiens misc_feature Incyte ID No 3143411CD1 15 Met Arg Pro Gly Thr Ala Leu Gln Ala Val Leu Leu Ala Val Leu 1 5 10 15 Leu Val Gly Leu Arg Ala Ala Thr Gly Arg Leu Leu Ser Gly Gln 20 25 30 Pro Val Cys Arg Gly Gly Thr Gln Arg Pro Cys Tyr Lys Val Ile 35 40 45 Tyr Phe His Asp Thr Ser Arg Arg Leu Asn Phe Glu Glu Ala Lys 50 55 60 Glu Ala Cys Arg Arg Asp Gly Gly Gln Leu Val Ser Ile Glu Ser 65 70 75 Glu Asp Glu Gln Lys Leu Ile Glu Lys Phe Ile Glu Asn Leu Leu 80 85 90 Pro Ser Asp Gly Asp Phe Trp Ile Gly Leu Arg Arg Arg Glu Glu 95 100 105 Lys Gln Ser Asn Ser Thr Ala Cys Gln Asp Leu Tyr Ala Trp Thr 110 115 120 Asp Gly Ser Ile Ser Gln Phe Arg Asn Trp Tyr Val Asp Glu Pro 125 130 135 Ser Cys Gly Ser Glu Val Cys Val Val Met Tyr His Gln Pro Ser 140 145 150 Ala Pro Ala Gly Ile Gly Gly Pro Tyr Met Phe Gln Trp Asn Asp 155 160 165 Asp Arg Cys Asn Met Lys Asn Asn Phe Ile Cys Lys Tyr Ser Asp 170 175 180 Glu Lys Pro Ala Val Pro Ser Arg Glu Ala Glu Gly Glu Glu Thr 185 190 195 Glu Leu Thr Thr Pro Val Leu Pro Glu Glu Thr Gln Glu Glu Asp 200 205 210 Ala Lys Lys Thr Phe Lys Glu Ser Arg Glu Ala Ala Leu Asn Leu 215 220 225 Ala Tyr Ile Leu Ile Pro Ser Ile Pro Leu Leu Leu Leu Leu Val 230 235 240 Val Thr Thr Val Val Cys Trp Val Trp Ile Cys Arg Lys Arg Lys 245 250 255 Arg Glu Gln Pro Asp Pro Ser Thr Lys Lys Gln His Thr Ile Trp 260 265 270 Pro Ser Pro His Gln Gly Asn Ser Pro Asp Leu Glu Val Tyr Asn 275 280 285 Val Ile Arg Lys Gln Ser Glu Ala Asp Leu Ala Glu Thr Arg Pro 290 295 300 Asp Leu Lys Asn Ile Ser Phe Arg Val Cys Ser Gly Glu Ala Thr 305 310 315 Pro Asp Asp Met Ser Cys Asp Tyr Asp Asn Met Ala Val Asn Pro 320 325 330 Ser Glu Ser Gly Phe Val Thr Leu Val Ser Val Glu Ser Gly Phe 335 340 345 Val Thr Asn Asp Ile Tyr Glu Phe Ser Pro Asp Gln Met Gly Arg 350 355 360 Ser Lys Glu Ser Gly Trp Val Glu Asn Glu Ile Tyr Gly Tyr 365 370 16 102 PRT Homo sapiens misc_feature Incyte ID No 3170835CD1 16 Met Lys Phe Ala Ile Val Leu Phe Ala Leu Phe Ala Val Ala Leu 1 5 10 15 Ala Ala Pro Thr Val Glu Val Leu Arg Ser Asp Ser Asn Val Gly 20 25 30 Ile Asp Asn Tyr Ser Tyr Ala Val Glu Thr Ser Asp Gly Thr Ser 35 40 45 Lys Ser Glu Glu Gly Val Leu Lys Asn Ala Gly Thr Glu Leu Glu 50 55 60 Ala Ile Ser Thr His Gly Ser Phe Ser Tyr Val Gly Pro Asp Gly 65 70 75 Gln Thr Tyr Thr Val Thr Tyr Val Ala Asp Glu Asn Gly Phe Gln 80 85 90 Pro Gln Gly Ala His Leu Pro Val Ala Pro Val Ala 95 100 17 510 PRT Homo sapiens misc_feature Incyte ID No 3550808CD1 17 Met Arg Pro Gly Leu Ser Phe Leu Leu Ala Leu Leu Phe Phe Leu 1 5 10 15 Gly Gln Ala Ala Gly Asp Leu Gly Asp Val Gly Pro Pro Ile Pro 20 25 30 Ser Pro Gly Phe Ser Ser Phe Pro Gly Val Asp Ser Ser Ser Ser 35 40 45 Phe Ser Ser Ser Ser Arg Ser Gly Ser Ser Ser Ser Arg Ser Leu 50 55 60 Gly Ser Gly Gly Ser Val Ser Gln Leu Phe Ser Asn Phe Thr Gly 65 70 75 Ser Val Asp Asp Arg Gly Thr Cys Gln Cys Ser Val Ser Leu Pro 80 85 90 Asp Thr Thr Phe Pro Val Asp Arg Val Glu Arg Leu Glu Phe Thr 95 100 105 Ala His Val Leu Ser Gln Lys Phe Glu Lys Glu Leu Ser Lys Val 110 115 120 Arg Glu Tyr Val Gln Leu Ile Ser Val Tyr Glu Lys Lys Leu Leu 125 130 135 Asn Leu Thr Val Arg Ile Asp Ile Met Glu Lys Asp Thr Ile Ser 140 145 150 Tyr Thr Glu Leu Asp Phe Glu Leu Ile Lys Val Glu Val Lys Glu 155 160 165 Met Glu Lys Leu Val Ile Gln Leu Lys Glu Ser Phe Gly Gly Ser 170 175 180 Ser Glu Ile Val Asp Gln Leu Glu Val Glu Ile Arg Asn Met Thr 185 190 195 Leu Leu Val Glu Lys Leu Glu Thr Leu Asp Lys Asn Asn Val Leu 200 205 210 Ala Ile Arg Arg Glu Ile Val Ala Leu Lys Thr Lys Leu Lys Glu 215 220 225 Cys Glu Ala Ser Lys Asp Gln Asn Thr Pro Val Val His Pro Pro 230 235 240 Pro Thr Pro Gly Ser Cys Gly His Gly Gly Val Val Asn Ile Ser 245 250 255 Lys Pro Ser Val Val Gln Leu Asn Trp Arg Gly Phe Ser Tyr Leu 260 265 270 Tyr Gly Ala Trp Gly Arg Asp Tyr Ser Pro Gln His Pro Asn Lys 275 280 285 Gly Leu Tyr Trp Val Ala Pro Leu Asn Thr Asp Gly Arg Leu Leu 290 295 300 Glu Tyr Tyr Arg Leu Tyr Asn Thr Leu Asp Asp Leu Leu Leu Tyr 305 310 315 Ile Asn Ala Arg Glu Leu Arg Ile Thr Tyr Gly Gln Gly Ser Gly 320 325 330 Thr Ala Val Tyr Asn Asn Asn Met Tyr Val Asn Met Tyr Asn Thr 335 340 345 Gly Asn Ile Ala Arg Val Asn Leu Thr Thr Asn Thr Ile Ala Val 350 355 360 Thr Gln Thr Leu Pro Asn Ala Ala Tyr Asn Asn Arg Phe Ser Tyr 365 370 375 Ala Asn Val Ala Trp Gln Asp Ile Asp Phe Ala Val Asp Glu Asn 380 385 390 Gly Leu Trp Val Ile Tyr Ser Thr Glu Ala Ser Thr Gly Asn Met 395 400 405 Val Ile Ser Lys Leu Asn Asp Thr Thr Leu Gln Val Leu Asn Thr 410 415 420 Trp Tyr Thr Lys Gln Tyr Lys Pro Ser Ala Ser Asn Ala Phe Met 425 430 435 Val Cys Gly Val Leu Tyr Ala Thr Arg Thr Met Asn Thr Arg Thr 440 445 450 Glu Glu Ile Phe Tyr Tyr Tyr Asp Thr Asn Thr Gly Lys Glu Gly 455 460 465 Lys Leu Asp Ile Val Met His Lys Met Gln Glu Lys Val Gln Ser 470 475 480 Ile Asn Tyr Asn Pro Phe Asp Gln Lys Leu Tyr Val Tyr Asn Asp 485 490 495 Gly Tyr Leu Leu Asn Tyr Asp Leu Ser Val Leu Gln Lys Pro Gln 500 505 510 18 185 PRT Homo sapiens misc_feature Incyte ID No 3683905CD1 18 Met Phe Leu Leu Asp Ser Ser Ala Ser Val Ser His Tyr Glu Phe 1 5 10 15 Ser Arg Val Arg Glu Phe Val Gly Gln Leu Val Ala Pro Leu Pro 20 25 30 Leu Gly Thr Gly Ala Leu Arg Ala Ser Leu Val His Val Gly Ser 35 40 45 Arg Pro Tyr Thr Glu Phe Pro Phe Gly Gln His Ser Ser Gly Glu 50 55 60 Ala Ala Gln Asp Ala Val Arg Ala Ser Ala Gln Arg Met Gly Asp 65 70 75 Thr His Thr Gly Leu Ala Leu Val Tyr Ala Lys Glu Gln Leu Phe 80 85 90 Ala Glu Ala Ser Gly Ala Arg Pro Gly Val Pro Lys Val Leu Val 95 100 105 Trp Val Thr Asp Gly Gly Ser Ser Asp Pro Val Gly Pro Pro Met 110 115 120 Gln Glu Leu Lys Asp Leu Gly Val Thr Val Phe Ile Val Ser Thr 125 130 135 Gly Arg Gly Asn Phe Leu Glu Leu Ser Ala Ala Ala Ser Ala Pro 140 145 150 Ala Glu Lys His Leu His Phe Val Asp Val Asp Asp Leu His Ile 155 160 165 Ile Val Gln Glu Leu Arg Gly Ser Ile Leu Asp Ala Met Arg Pro 170 175 180 Gln Ala Tyr Ser Leu 185 19 207 PRT Homo sapiens misc_feature Incyte ID No 4062841CD1 19 Met Ala Ala Leu Val Glu Pro Leu Gly Leu Glu Arg Asp Val Ser 1 5 10 15 Arg Ala Val Glu Leu Leu Glu Arg Leu Gln Arg Ser Gly Glu Leu 20 25 30 Pro Pro Gln Lys Leu Gln Ala Leu Gln Arg Val Leu Gln Ser Arg 35 40 45 Phe Cys Ser Ala Ile Arg Glu Val Tyr Glu Gln Leu Tyr Asp Thr 50 55 60 Leu Asp Ile Thr Gly Ser Ala Glu Ile Arg Ala His Ala Thr Ala 65 70 75 Lys Ala Thr Val Ala Ala Phe Thr Ala Ser Glu Gly His Ala His 80 85 90 Pro Arg Val Val Glu Leu Pro Lys Thr Asp Glu Gly Leu Gly Phe 95 100 105 Asn Ile Met Gly Gly Lys Glu Gln Asn Ser Pro Ile Tyr Ile Ser 110 115 120 Arg Val Ile Pro Gly Gly Val Ala Asp Arg His Gly Gly Leu Lys 125 130 135 Arg Gly Asp Gln Leu Leu Ser Val Asn Gly Val Ser Val Glu Gly 140 145 150 Glu Gln His Glu Lys Ala Val Glu Leu Leu Lys Ala Ala Gln Gly 155 160 165 Ser Val Lys Leu Val Val Arg Tyr Thr Pro Arg Val Leu Glu Glu 170 175 180 Met Glu Ala Arg Phe Glu Lys Met Arg Ser Ala Arg Arg Arg Gln 185 190 195 Gln His Gln Ser Tyr Ser Ser Leu Glu Ser Arg Gly 200 205 20 238 PRT Homo sapiens misc_feature Incyte ID No 6394358CD1 20 Met Ser Leu Asn Glu His Ser Met Gln Ala Leu Ser Trp Arg Lys 1 5 10 15 Leu Tyr Leu Ser Arg Ala Lys Leu Lys Ala Ser Ser Arg Thr Ser 20 25 30 Ala Leu Leu Ser Gly Phe Ala Met Val Ala Met Val Glu Val Gln 35 40 45 Leu Asp Ala Asp His Asp Tyr Pro Pro Gly Leu Leu Ile Ala Phe 50 55 60 Ser Ala Cys Thr Thr Val Leu Val Ala Val His Leu Phe Ala Leu 65 70 75 Met Ile Ser Thr Cys Ile Leu Pro Asn Ile Glu Ala Val Ser Asn 80 85 90 Val His Asn Leu Asn Ser Val Lys Glu Ser Pro His Glu Arg Met 95 100 105 His Arg His Ile Glu Leu Ala Trp Ala Phe Ser Thr Val Ile Gly 110 115 120 Thr Leu Leu Phe Leu Ala Glu Val Val Leu Leu Cys Trp Val Lys 125 130 135 Phe Leu Pro Leu Lys Lys Gln Pro Gly Gln Pro Arg Pro Thr Ser 140 145 150 Lys Pro Pro Ala Ser Gly Ala Ala Ala Asn Val Ser Thr Ser Gly 155 160 165 Ile Thr Pro Gly Gln Ala Ala Ala Ile Ala Ser Thr Thr Ile Met 170 175 180 Val Pro Phe Gly Leu Ile Phe Ile Val Phe Ala Val His Phe Tyr 185 190 195 Arg Ser Leu Val Ser His Lys Thr Asp Arg Gln Phe Gln Glu Leu 200 205 210 Asn Glu Leu Ala Glu Phe Ala Arg Leu Gln Asp Gln Leu Asp His 215 220 225 Arg Gly Asp His Pro Leu Thr Pro Gly Ser His Tyr Ala 230 235 21 3298 PRT Homo sapiens misc_feature Incyte ID No 2847752CD1 21 Met Ala Arg Arg Pro Pro Trp Arg Gly Leu Gly Gly Arg Ser Thr 1 5 10 15 Pro Ile Leu Leu Leu Leu Leu Leu Ser Leu Phe Pro Leu Ser Gln 20 25 30 Glu Glu Leu Gly Gly Gly Gly His Gln Gly Trp Asp Pro Gly Leu 35 40 45 Ala Ala Thr Thr Gly Pro Arg Ala His Ile Gly Gly Gly Ala Leu 50 55 60 Ala Leu Cys Pro Glu Ser Ser Gly Val Arg Glu Asp Gly Gly Pro 65 70 75 Gly Leu Gly Val Arg Glu Pro Ile Phe Val Gly Leu Arg Gly Arg 80 85 90 Arg Gln Ser Ala Arg Asn Ser Arg Gly Pro Pro Glu Gln Pro Asn 95 100 105 Glu Glu Leu Gly Ile Glu His Gly Val Gln Pro Leu Gly Ser Arg 110 115 120 Glu Arg Glu Thr Gly Gln Gly Pro Gly Ser Val Leu Tyr Trp Arg 125 130 135 Pro Glu Val Ser Ser Cys Gly Arg Thr Gly Pro Leu Gln Arg Gly 140 145 150 Ser Leu Ser Pro Gly Ala Leu Ser Ser Gly Val Pro Gly Ser Gly 155 160 165 Asn Ser Ser Pro Leu Pro Ser Asp Phe Leu Ile Arg His His Gly 170 175 180 Pro Lys Pro Val Ser Ser Gln Arg Asn Ala Gly Thr Gly Ser Arg 185 190 195 Lys Arg Val Gly Thr Ala Arg Cys Cys Gly Glu Leu Trp Ala Thr 200 205 210 Gly Ser Lys Gly Gln Gly Glu Arg Ala Thr Thr Ser Gly Ala Glu 215 220 225 Arg Thr Ala Pro Arg Arg Asn Cys Leu Pro Gly Ala Ser Gly Ser 230 235 240 Gly Pro Glu Leu Asp Ser Ala Pro Arg Thr Ala Arg Thr Ala Pro 245 250 255 Ala Ser Gly Ser Ala Pro Arg Glu Ser Arg Thr Ala Pro Glu Pro 260 265 270 Ala Pro Lys Arg Met Arg Ser Arg Gly Leu Phe Arg Cys Arg Phe 275 280 285 Leu Pro Gln Arg Pro Gly Pro Arg Pro Pro Gly Leu Pro Ala Arg 290 295 300 Pro Glu Ala Arg Lys Val Thr Ser Ala Asn Arg Ala Arg Phe Arg 305 310 315 Arg Ala Ala Asn Arg His Pro Gln Phe Pro Gln Tyr Asn Tyr Gln 320 325 330 Thr Leu Val Pro Glu Asn Glu Ala Ala Gly Thr Ala Val Leu Arg 335 340 345 Val Val Ala Gln Asp Pro Asp Ala Gly Glu Ala Gly Arg Leu Val 350 355 360 Tyr Ser Leu Ala Ala Leu Met Asn Ser Arg Ser Leu Glu Leu Phe 365 370 375 Ser Ile Asp Pro Gln Ser Gly Leu Ile Arg Thr Ala Ala Ala Leu 380 385 390 Asp Arg Glu Ser Met Glu Arg His Tyr Leu Arg Val Thr Ala Gln 395 400 405 Asp His Gly Ser Pro Arg Leu Ser Ala Thr Thr Met Val Ala Val 410 415 420 Thr Val Ala Asp Arg Asn Asp His Ser Pro Val Phe Glu Gln Ala 425 430 435 Gln Tyr Arg Glu Thr Leu Arg Glu Asn Val Glu Glu Gly Tyr Pro 440 445 450 Ile Leu Gln Leu Arg Ala Thr Asp Gly Asp Ala Pro Pro Asn Ala 455 460 465 Asn Leu Arg Tyr Arg Phe Val Gly Pro Pro Ala Ala Arg Ala Ala 470 475 480 Ala Ala Ala Ala Phe Glu Ile Asp Pro Arg Ser Gly Leu Ile Ser 485 490 495 Thr Ser Gly Arg Val Asp Arg Glu His Met Glu Ser Tyr Glu Leu 500 505 510 Val Val Glu Ala Ser Asp Gln Gly Gln Glu Pro Gly Pro Arg Ser 515 520 525 Ala Thr Val Arg Val His Ile Thr Val Leu Asp Glu Asn Asp Asn 530 535 540 Ala Pro Gln Phe Ser Glu Lys Arg Tyr Val Ala Gln Val Arg Glu 545 550 555 Asp Val Arg Pro His Thr Val Val Leu Arg Val Thr Ala Thr Asp 560 565 570 Arg Asp Lys Asp Ala Asn Gly Leu Val His Tyr Asn Ile Ile Ser 575 580 585 Gly Asn Ser Arg Gly His Phe Ala Ile Asp Ser Leu Thr Gly Glu 590 595 600 Ile Gln Val Val Ala Pro Leu Asp Phe Glu Ala Glu Arg Glu Tyr 605 610 615 Ala Leu Arg Ile Arg Ala Gln Asp Ala Gly Arg Pro Pro Leu Ser 620 625 630 Asn Asn Thr Gly Leu Ala Ser Ile Gln Val Val Asp Ile Asn Asp 635 640 645 His Ile Pro Ile Phe Val Ser Thr Pro Phe Gln Val Ser Val Leu 650 655 660 Glu Asn Ala Pro Leu Gly His Ser Val Ile His Ile Gln Ala Val 665 670 675 Asp Ala Asp His Gly Glu Asn Ala Arg Leu Glu Tyr Ser Leu Thr 680 685 690 Gly Val Ala Pro Asp Thr Pro Phe Val Ile Asn Ser Ala Thr Gly 695 700 705 Trp Val Ser Val Ser Gly Pro Leu Asp Arg Glu Ser Val Glu His 710 715 720 Tyr Phe Phe Gly Val Glu Ala Arg Asp His Gly Ser Pro Pro Leu 725 730 735 Ser Ala Ser Ala Ser Val Thr Val Thr Val Leu Asp Val Asn Asp 740 745 750 Asn Arg Pro Glu Phe Thr Met Lys Glu Tyr His Leu Arg Leu Asn 755 760 765 Glu Asp Ala Ala Val Gly Thr Ser Val Val Ser Val Thr Ala Val 770 775 780 Asp Arg Asp Ala Asn Ser Ala Ile Ser Tyr Gln Ile Thr Gly Gly 785 790 795 Asn Thr Arg Asn Arg Phe Ala Ile Ser Thr Gln Gly Gly Val Gly 800 805 810 Leu Val Thr Leu Ala Leu Pro Leu Asp Tyr Lys Gln Glu Arg Tyr 815 820 825 Phe Lys Leu Val Leu Thr Ala Ser Asp Arg Ala Leu His Asp His 830 835 840 Cys Tyr Val His Ile Asn Ile Thr Asp Ala Asn Thr His Arg Pro 845 850 855 Val Phe Gln Ser Ala His Tyr Ser Val Ser Val Asn Glu Asp Arg 860 865 870 Pro Met Gly Ser Thr Ile Val Val Ile Ser Ala Ser Asp Asp Asp 875 880 885 Val Gly Glu Asn Ala Arg Ile Thr Tyr Leu Leu Glu Asp Asn Leu 890 895 900 Pro Gln Phe Arg Ile Asp Ala Asp Ser Gly Ala Ile Thr Leu Gln 905 910 915 Ala Pro Leu Asp Tyr Glu Asp Gln Val Thr Tyr Thr Leu Ala Ile 920 925 930 Thr Ala Arg Asp Asn Gly Ile Pro Gln Lys Ala Asp Thr Thr Tyr 935 940 945 Val Glu Val Met Val Asn Asp Val Asn Asp Asn Ala Pro Gln Phe 950 955 960 Val Ala Ser His Tyr Thr Gly Leu Val Ser Glu Asp Ala Pro Pro 965 970 975 Phe Thr Ser Val Leu Gln Ile Ser Ala Thr Asp Arg Asp Ala His 980 985 990 Ala Asn Gly Arg Val Gln Tyr Thr Phe Gln Asn Gly Glu Asp Gly 995 1000 1005 Asp Gly Asp Phe Thr Ile Glu Pro Thr Ser Gly Ile Val Arg Thr 1010 1015 1020 Val Arg Arg Leu Asp Arg Glu Ala Val Ser Val Tyr Glu Leu Thr 1025 1030 1035 Ala Tyr Ala Val Asp Arg Gly Val Pro Pro Leu Arg Thr Pro Val 1040 1045 1050 Ser Ile Gln Val Met Val Gln Asp Val Asn Asp Asn Ala Pro Val 1055 1060 1065 Phe Pro Ala Glu Glu Phe Glu Val Arg Val Lys Glu Asn Ser Ile 1070 1075 1080 Val Gly Ser Val Val Ala Gln Ile Thr Ala Val Asp Pro Asp Glu 1085 1090 1095 Gly Pro Asn Ala His Ile Met Tyr Gln Ile Val Glu Gly Asn Ile 1100 1105 1110 Pro Glu Leu Phe Gln Met Asp Ile Phe Ser Gly Glu Leu Thr Ala 1115 1120 1125 Leu Ile Asp Leu Asp Tyr Glu Ala Arg Gln Glu Tyr Val Ile Val 1130 1135 1140 Val Gln Ala Thr Ser Ala Pro Leu Val Ser Arg Ala Thr Val His 1145 1150 1155 Val Arg Leu Val Asp Gln Asn Asp Asn Ser Pro Val Leu Asn Asn 1160 1165 1170 Phe Gln Ile Leu Phe Asn Asn Tyr Val Ser Asn Arg Ser Asp Thr 1175 1180 1185 Phe Pro Ser Gly Ile Ile Gly Arg Ile Pro Ala Tyr Asp Pro Asp 1190 1195 1200 Val Ser Asp His Leu Phe Tyr Ser Phe Glu Arg Gly Asn Glu Leu 1205 1210 1215 Gln Leu Leu Val Val Asn Gln Thr Ser Gly Glu Leu Arg Leu Ser 1220 1225 1230 Arg Lys Leu Asp Asn Asn Arg Pro Leu Val Ala Ser Met Leu Val 1235 1240 1245 Thr Val Thr Asp Gly Leu His Ser Val Thr Ala Gln Cys Val Leu 1250 1255 1260 Arg Val Val Ile Ile Thr Glu Glu Leu Leu Ala Asn Ser Leu Thr 1265 1270 1275 Val Arg Leu Glu Asn Met Trp Gln Glu Arg Phe Leu Ser Pro Leu 1280 1285 1290 Leu Gly Arg Phe Leu Glu Gly Val Ala Ala Val Leu Ala Thr Pro 1295 1300 1305 Ala Glu Asp Val Phe Ile Phe Asn Ile Gln Asn Asp Thr Asp Val 1310 1315 1320 Gly Gly Thr Val Leu Asn Val Ser Phe Ser Ala Leu Ala Pro Arg 1325 1330 1335 Gly Ala Gly Ala Gly Ala Ala Gly Pro Trp Phe Ser Ser Glu Glu 1340 1345 1350 Leu Gln Glu Gln Leu Tyr Val Arg Arg Ala Ala Leu Ala Ala Arg 1355 1360 1365 Ser Leu Leu Asp Val Leu Pro Phe Asp Asp Asn Val Cys Leu Arg 1370 1375 1380 Glu Pro Cys Glu Asn Tyr Met Lys Cys Val Ser Val Leu Arg Phe 1385 1390 1395 Asp Ser Ser Ala Pro Phe Leu Ala Ser Ala Ser Thr Leu Phe Arg 1400 1405 1410 Pro Ile Gln Pro Ile Ala Gly Leu Arg Cys Arg Cys Pro Pro Gly 1415 1420 1425 Phe Thr Gly Asp Phe Cys Glu Thr Glu Leu Asp Leu Cys Tyr Ser 1430 1435 1440 Asn Pro Cys Arg Asn Gly Gly Ala Cys Ala Arg Arg Glu Gly Gly 1445 1450 1455 Tyr Thr Cys Val Cys Arg Pro Arg Phe Thr Gly Glu Asp Cys Glu 1460 1465 1470 Leu Asp Thr Glu Ala Gly Arg Cys Val Pro Gly Val Cys Arg Asn 1475 1480 1485 Gly Gly Thr Cys Thr Asp Ala Pro Asn Gly Gly Phe Arg Cys Gln 1490 1495 1500 Cys Pro Ala Gly Gly Ala Phe Glu Gly Pro Arg Cys Glu Val Ala 1505 1510 1515 Ala Arg Ser Phe Pro Pro Ser Ser Phe Val Met Phe Arg Gly Leu 1520 1525 1530 Arg Gln Arg Phe His Leu Thr Leu Ser Leu Ser Phe Ala Thr Val 1535 1540 1545 Gln Gln Ser Gly Leu Leu Phe Tyr Asn Gly Arg Leu Asn Glu Lys 1550 1555 1560 His Asp Phe Leu Ala Leu Glu Leu Val Ala Gly Gln Val Arg Leu 1565 1570 1575 Thr Tyr Ser Thr Gly Glu Ser Asn Thr Val Val Ser Pro Thr Val 1580 1585 1590 Pro Gly Gly Leu Ser Asp Gly Gln Trp His Thr Val His Leu Arg 1595 1600 1605 Tyr Tyr Asn Lys Pro Arg Thr Asp Ala Leu Gly Gly Ala Gln Gly 1610 1615 1620 Pro Ser Lys Asp Lys Val Ala Val Leu Ser Val Asp Asp Cys Asp 1625 1630 1635 Val Ala Val Ala Leu Gln Phe Gly Ala Glu Ile Gly Asn Tyr Ser 1640 1645 1650 Cys Ala Ala Ala Gly Val Gln Thr Ser Ser Lys Lys Ser Leu Asp 1655 1660 1665 Leu Thr Gly Pro Leu Leu Leu Gly Gly Val Pro Asn Leu Pro Glu 1670 1675 1680 Asn Phe Pro Val Ser His Lys Asp Phe Ile Gly Cys Met Arg Asp 1685 1690 1695 Leu His Ile Asp Gly Arg Arg Val Asp Met Ala Ala Phe Val Ala 1700 1705 1710 Asn Asn Gly Thr Met Ala Gly Cys Gln Ala Lys Leu His Phe Cys 1715 1720 1725 Asp Ser Gly Pro Cys Lys Asn Ser Gly Phe Cys Ser Glu Arg Trp 1730 1735 1740 Gly Ser Phe Ser Cys Asp Cys Pro Val Gly Phe Gly Gly Lys Asp 1745 1750 1755 Cys Gln Leu Thr Met Ala His Pro His His Phe Arg Gly Asn Gly 1760 1765 1770 Thr Leu Ser Trp Asn Phe Gly Ser Asp Met Ala Val Ser Val Pro 1775 1780 1785 Trp Tyr Leu Gly Leu Ala Phe Arg Thr Arg Ala Thr Gln Gly Val 1790 1795 1800 Leu Met Gln Val Gln Ala Gly Pro His Ser Thr Leu Leu Cys Gln 1805 1810 1815 Leu Asp Arg Gly Leu Leu Ser Val Thr Val Thr Arg Gly Ser Gly 1820 1825 1830 Arg Ala Ser His Leu Leu Leu Asp Gln Val Thr Val Ser Asp Gly 1835 1840 1845 Arg Trp His Asp Leu Arg Leu Glu Leu Gln Glu Glu Pro Gly Gly 1850 1855 1860 Arg Arg Gly His His Val Leu Met Val Ser Leu Asp Phe Ser Leu 1865 1870 1875 Phe Gln Asp Thr Met Ala Val Gly Ser Glu Leu Gln Gly Leu Lys 1880 1885 1890 Val Lys Gln Leu His Val Gly Gly Leu Pro Pro Gly Ser Ala Glu 1895 1900 1905 Glu Ala Pro Gln Gly Leu Val Gly Cys Ile Gln Pro Pro Ser Glu 1910 1915 1920 Cys Gly Pro Gly Cys Val Val Thr Asn Ala Cys Ala Ser Gly Pro 1925 1930 1935 Cys Pro Pro His Ala Asp Cys Arg Asp Leu Trp Gln Thr Phe Ser 1940 1945 1950 Cys Thr Cys Gln Pro Gly Tyr Tyr Gly Pro Gly Cys Val Asp Ala 1955 1960 1965 Cys Leu Leu Asn Pro Cys Gln Asn Gln Gly Ser Cys Arg His Leu 1970 1975 1980 Pro Gly Ala Pro His Gly Tyr Thr Cys Asp Cys Val Gly Gly Tyr 1985 1990 1995 Phe Gly His His Cys Glu His Arg Met Asp Gln Gln Cys Pro Arg 2000 2005 2010 Gly Trp Trp Gly Ser Pro Thr Cys Gly Pro Cys Asn Cys Asp Val 2015 2020 2025 His Lys Gly Phe Asp Pro Asn Cys Asn Lys Thr Asn Gly Gln Cys 2030 2035 2040 His Cys Lys Glu Phe His Tyr Arg Pro Arg Gly Ser Asp Ser Cys 2045 2050 2055 Leu Pro Cys Asp Cys Tyr Pro Val Gly Ser Thr Ser Arg Ser Cys 2060 2065 2070 Ala Pro His Ser Gly Gln Cys Pro Cys Arg Pro Gly Ala Leu Gly 2075 2080 2085 Arg Gln Cys Asn Ser Cys Asp Ser Pro Phe Ala Glu Val Thr Ala 2090 2095 2100 Ser Gly Cys Arg Val Leu Tyr Asp Ala Cys Pro Lys Ser Leu Arg 2105 2110 2115 Ser Gly Val Trp Trp Pro Gln Thr Lys Phe Gly Val Leu Ala Thr 2120 2125 2130 Val Pro Cys Pro Arg Gly Ala Leu Gly Leu Arg Gly Ala Gly Ala 2135 2140 2145 Ala Val Arg Leu Cys Asp Glu Ala Gln Gly Trp Leu Glu Pro Asp 2150 2155 2160 Leu Phe Asn Cys Thr Ser Pro Ala Phe Arg Glu Leu Ser Leu Leu 2165 2170 2175 Leu Asp Gly Leu Glu Leu Asn Lys Thr Ala Leu Asp Thr Met Glu 2180 2185 2190 Ala Lys Lys Leu Ala Gln Arg Leu Arg Glu Val Thr Gly His Thr 2195 2200 2205 Asp His Tyr Phe Ser Gln Asp Val Arg Val Thr Ala Arg Leu Leu 2210 2215 2220 Ala His Leu Leu Ala Phe Glu Ser His Gln Gln Gly Phe Gly Leu 2225 2230 2235 Thr Ala Thr Gln Asp Ala His Phe Asn Glu Asn Leu Leu Trp Ala 2240 2245 2250 Gly Ser Ala Leu Leu Ala Pro Glu Thr Gly Asp Leu Trp Ala Ala 2255 2260 2265 Leu Gly Gln Arg Ala Pro Gly Gly Ser Pro Gly Ser Ala Gly Leu 2270 2275 2280 Val Arg His Leu Glu Glu Tyr Ala Ala Thr Leu Ala Arg Asn Met 2285 2290 2295 Glu Leu Thr Tyr Leu Asn Pro Met Gly Leu Val Thr Pro Asn Ile 2300 2305 2310 Met Leu Ser Ile Asp Arg Met Glu His Pro Ser Ser Pro Arg Gly 2315 2320 2325 Ala Arg Arg Tyr Pro Arg Tyr His Ser Asn Leu Phe Arg Gly Gln 2330 2335 2340 Asp Ala Trp Asp Pro His Thr His Val Leu Leu Pro Ser Gln Ser 2345 2350 2355 Pro Arg Pro Ser Pro Ser Glu Val Leu Pro Thr Ser Ser Ser Ile 2360 2365 2370 Glu Asn Ser Thr Thr Ser Ser Val Val Pro Pro Pro Ala Pro Pro 2375 2380 2385 Glu Pro Glu Pro Gly Ile Ser Ile Ile Ile Leu Leu Val Tyr Arg 2390 2395 2400 Thr Leu Gly Gly Leu Leu Pro Ala Gln Phe Gln Ala Glu Arg Arg 2405 2410 2415 Gly Ala Arg Leu Pro Gln Asn Pro Val Met Asn Ser Pro Val Val 2420 2425 2430 Ser Val Ala Val Phe His Gly Arg Asn Phe Leu Arg Gly Ile Leu 2435 2440 2445 Glu Ser Pro Ile Ser Leu Glu Phe Arg Leu Leu Gln Thr Ala Asn 2450 2455 2460 Arg Ser Lys Ala Ile Cys Val Gln Trp Asp Pro Pro Gly Leu Ala 2465 2470 2475 Glu Gln His Gly Val Trp Thr Ala Arg Asp Cys Glu Leu Val His 2480 2485 2490 Arg Asn Gly Ser His Ala Arg Cys Arg Cys Ser Arg Thr Gly Thr 2495 2500 2505 Phe Gly Val Leu Met Asp Ala Ser Pro Arg Glu Arg Leu Glu Gly 2510 2515 2520 Asp Leu Glu Leu Leu Ala Val Phe Thr His Val Val Val Ala Val 2525 2530 2535 Ser Val Ala Ala Leu Val Leu Thr Ala Ala Ile Leu Leu Ser Leu 2540 2545 2550 Arg Ser Leu Lys Ser Asn Val Arg Gly Ile His Ala Asn Val Ala 2555 2560 2565 Ala Ala Leu Gly Val Ala Glu Leu Leu Phe Leu Leu Gly Ile His 2570 2575 2580 Arg Thr His Asn Gln Leu Val Cys Thr Ala Val Ala Ile Leu Leu 2585 2590 2595 His Tyr Phe Phe Leu Ser Thr Phe Ala Trp Leu Phe Val Gln Gly 2600 2605 2610 Leu His Leu Tyr Arg Met Gln Val Glu Pro Arg Asn Val Asp Arg 2615 2620 2625 Gly Ala Met Arg Phe Tyr His Ala Leu Gly Trp Gly Val Pro Ala 2630 2635 2640 Val Leu Leu Gly Leu Ala Val Gly Leu Asp Pro Glu Gly Tyr Gly 2645 2650 2655 Asn Pro Asp Phe Cys Trp Ile Ser Val His Glu Pro Leu Ile Trp 2660 2665 2670 Ser Phe Ala Gly Pro Val Val Leu Val Ile Val Met Asn Gly Thr 2675 2680 2685 Met Phe Leu Leu Ala Ala Arg Thr Ser Cys Ser Thr Gly Gln Arg 2690 2695 2700 Glu Ala Lys Lys Thr Ser Ala Leu Thr Leu Arg Ser Ser Phe Leu 2705 2710 2715 Leu Leu Leu Leu Val Ser Ala Ser Trp Leu Phe Gly Leu Leu Ala 2720 2725 2730 Val Asn His Ser Ile Leu Ala Phe His Tyr Leu His Ala Gly Leu 2735 2740 2745 Cys Gly Leu Gln Gly Leu Ala Val Leu Leu Leu Phe Cys Val Leu 2750 2755 2760 Asn Ala Asp Ala Arg Ala Ala Trp Met Pro Ala Cys Leu Gly Arg 2765 2770 2775 Lys Ala Ala Pro Glu Glu Ala Arg Pro Ala Pro Gly Leu Gly Pro 2780 2785 2790 Gly Ala Tyr Asn Asn Thr Ala Leu Phe Glu Glu Ser Gly Leu Ile 2795 2800 2805 Arg Ile Thr Leu Gly Ala Ser Thr Val Ser Ser Val Ser Ser Ala 2810 2815 2820 Arg Ser Gly Arg Thr Gln Asp Gln Asp Ser Gln Arg Gly Arg Ser 2825 2830 2835 Tyr Leu Arg Asp Asn Val Leu Val Arg His Gly Ser Ala Ala Asp 2840 2845 2850 His Thr Asp His Ser Leu Gln Ala His Ala Gly Pro Thr Asp Leu 2855 2860 2865 Asp Val Ala Met Phe His Arg Asp Ala Gly Ala Asp Ser Asp Ser 2870 2875 2880 Asp Ser Asp Leu Ser Leu Glu Glu Glu Arg Ser Leu Ser Ile Pro 2885 2890 2895 Ser Ser Glu Ser Glu Asp Asn Gly Arg Thr Arg Gly Arg Val Gln 2900 2905 2910 Arg Pro Leu Cys Arg Ala Ala Gln Ser Glu Arg Leu Leu Thr His 2915 2920 2925 Pro Lys Asp Val Asp Gly Asn Asp Leu Leu Ser Tyr Trp Pro Ala 2930 2935 2940 Leu Gly Glu Cys Glu Ala Ala Pro Cys Ala Leu Gln Thr Trp Gly 2945 2950 2955 Ser Glu Arg Arg Leu Gly Leu Asp Thr Ser Lys Asp Ala Ala Asn 2960 2965 2970 Asn Asn Gln Pro Asp Pro Ala Leu Thr Ser Gly Asp Glu Thr Ser 2975 2980 2985 Leu Gly Arg Ala Gln Arg Gln Arg Lys Gly Ile Leu Lys Asn Arg 2990 2995 3000 Leu Gln Tyr Pro Leu Val Pro Gln Thr Arg Gly Ala Pro Glu Leu 3005 3010 3015 Ser Trp Cys Arg Ala Ala Thr Leu Gly His Arg Ala Val Pro Ala 3020 3025 3030 Ala Ser Tyr Gly Arg Ile Tyr Ala Gly Gly Gly Thr Gly Ser Leu 3035 3040 3045 Ser Gln Pro Ala Ser Arg Tyr Ser Ser Arg Glu Gln Leu Asp Leu 3050 3055 3060 Leu Leu Arg Arg Gln Leu Ser Arg Glu Arg Leu Glu Glu Ala Pro 3065 3070 3075 Ala Pro Val Leu Arg Pro Leu Ser Arg Pro Gly Ser Gln Glu Cys 3080 3085 3090 Met Asp Ala Ala Pro Gly Arg Leu Glu Pro Lys Asp Arg Gly Ser 3095 3100 3105 Thr Leu Pro Arg Arg Gln Pro Pro Arg Asp Tyr Pro Gly Ala Met 3110 3115 3120 Ala Gly Arg Phe Gly Ser Arg Asp Ala Leu Asp Leu Gly Ala Pro 3125 3130 3135 Arg Glu Trp Leu Ser Thr Leu Pro Pro Pro Arg Arg Thr Arg Asp 3140 3145 3150 Leu Asp Pro Gln Pro Pro Pro Leu Pro Leu Ser Pro Gln Arg Gln 3155 3160 3165 Leu Ser Arg Asp Pro Leu Leu Pro Ser Arg Pro Leu Asp Ser Leu 3170 3175 3180 Ser Arg Ser Ser Asn Ser Arg Glu Gln Leu Asp Gln Val Pro Ser 3185 3190 3195 Arg His Pro Ser Arg Glu Ala Leu Gly Pro Leu Pro Gln Leu Leu 3200 3205 3210 Arg Ala Arg Glu Asp Ser Val Ser Gly Pro Ser His Gly Pro Ser 3215 3220 3225 Thr Glu Gln Leu Asp Ile Leu Ser Ser Ile Leu Ala Ser Phe Asn 3230 3235 3240 Ser Ser Ala Leu Ser Ser Val Gln Ser Ser Ser Thr Pro Leu Gly 3245 3250 3255 Pro His Thr Thr Ala Thr Pro Ser Ala Thr Ala Ser Val Leu Gly 3260 3265 3270 Pro Ser Thr Pro Arg Ser Ala Thr Ser His Ser Ile Ser Glu Leu 3275 3280 3285 Ser Pro Asp Ser Glu Val Pro Arg Ser Glu Gly His Ser 3290 3295 22 905 DNA Homo sapiens misc_feature Incyte ID No 1424691CB1 22 cccacgcgtc cgtgttagaa ctccaaaaag cactaaacat gtccattcga ttcagtggag 60 tagaacaaaa cctcaggatg aagtgaaagc agtccaactt gccattcaga cattattcac 120 caattcagat ggcaaccctg gaagcaggtc cgactcaagt gctgattgcc agtggttaga 180 tactctgagg atgcggcaga ttgcatccaa cacttcttta cagcgttccc agagcaatcc 240 tattctgggg tcaccgttct tctcacactt tgatggccag gattcctacg ctgctgctgt 300 gagacggccc caggtgccca ttaagtatca acagattaca cctgtgaacc agtccagaag 360 ctcgtctcct actcagtatg gactgaccaa aaacttctct tccctacatc tcaactctag 420 ggacagtggc ttttccagtg gcaatactga cacctcttca gagaggggtc gatactcaga 480 cagaagcagg aacaaatatg gacgtggtag tatatcactc aattcttctc ctagaggaag 540 atacagtgga aagagtcagc attccactcc ttcaagagga agataccctg gaaagttcta 600 cagggtttct cagtcagcac tcaatcctca ccagtcgcct gacttcaaga gaagccccag 660 ggacctccac caacccaaca ccataccagg gatgcctttg caccctgaga ctgactcaag 720 agccagtgaa gaggacagca aagtcagcga agggggctgg acaaaagtgg aataccggaa 780 aaagccccac aggccatctc ccgccaaaac caataaagag agagccagag gggaccaccg 840 tggatggaga aacttttgat gaattgaact acatagcttt tctaagcagg ttaaaaaaaa 900 aaaaa 905 23 2390 DNA Homo sapiens misc_feature Incyte ID No 1450801CB1 23 tgaccttggc accccgcttc ctgccaaacg ccctgatcag tgtggacttt gctgccaact 60 atctcaccaa gatctatggg ctcacctttg gccagaagcc aaacttgagg tctgtgtacc 120 tgcacaacaa caagctggca gacgccgggc tgccggacaa catgttcaac ggctccagca 180 acgtcgaggt cctcatcctg tccagcaact tcctgcgcca cgtgcccaag cacctgccgc 240 ctgccctgta caagctgcac ctcaagaaca acaagctgga gaagatcccc ccgggggcct 300 tgcagcgagc tggagcagcc tgcgcgagct atacctgcag aacaactacc tgactgacga 360 gggcctggac aacgagacct tctggaagct ctccagcctg gagtacctgg atctgtccag 420 caacaacctg tctcgggtcc cagctgggct gccgcgcagc ctggtgctgc tgcacttgga 480 gaagaacgcc atccggagcg tggacgcgaa tgtgctgacc cccatccgca gcctggagta 540 cctgctgctg cacagcaacc agctgcggga gcagggcatc cacccactgg ccttccaggg 600 cctcaagcgg ttgcacacgg tgcacctgta caacaacgcg ctggagcgcg tgcccagtgg 660 cctgcctcgc cgcgtgcgca ccctcatgat cctgcacaac cagatcacag gcattggccg 720 cgaagacttt gccaccacct acttcctgga ggagctcaac ctcagctaca accgcatcac 780 cagcccacag gtgcaccgcg acgccttccg caagctgcgc ctgctgcgct cgctggacct 840 gtcgggcaac cggctgcaca cgctgccacc tgggctgcct cgaaatgtcc atgtgctgaa 900 ggtcaagcgc aatgagctgg ctgccttggc acgaggggcg ctggcgggca tggctcagct 960 gcgtgagctg tacctcacca gcaaccgact gcgcagccga gccctgggcc cccgtgcctg 1020 ggtggacctc gcccatctgc agctgctgga catcgccggg aatcagctca cagagatccc 1080 cgaggggctc cccgagtcac ttgagtacct gtacctgcag aacaacaaga ttagtgcggt 1140 gcccgccaat gccttcgact ccacgcccaa cctcaagggg atctttctca ggtttaacaa 1200 gctggctgtg ggctccgtgg tggacagtgc cttccggagg ctgaagcacc tgcaggtctt 1260 ggacattgaa ggcaacttag agtttggtga catttccaag gaccgtggcc gcttggggaa 1320 ggaaaaggag gaggaggaag aggaggagga ggaggaagag gaaacaagat agtgacaagg 1380 tgatgcagat gtgacctagg atgatggacc gccggactct tttctgcagc acacgcctgt 1440 gtgctgtgag ccccccactc tgccgtgctc acacagacac acccagctgc acacatgagg 1500 catcccacat gacacgggct gacacagtct catatcccca ccccttccca cggcgtgtcc 1560 cacggccaga cacatgcaca cacatcacac cctcaaacac ccagctcagc cacacacaac 1620 taccctccaa accaccacag tctctgtcac acccccacta ccgctgccac gccctctgaa 1680 tcatgcaggg aagggtctgc ccctgccctg gcacacacag gcacccattc cctccccctg 1740 ctgacatgtg tatgcgtatg catacacacc acacacacac acatgcacaa gtcatgtgcg 1800 aacagccctc caaagcctat gccacagaca gctcttgccc cagccagaat cagccatagc 1860 agctcgccgt ctgccctgtc catctgtccg tccgttccct ggagaagaca caagggtatc 1920 catgctctgt ggccaggtgc ctgccaccct ctggaactca caaaagctgg cttttattcc 1980 tttcccatcc tatggggaca ggagccttca ggactgctgg cctggcctgg cccaccctgc 2040 tcctccaggt gctgggcagt cactctgcta agagtccctc cctgccacgc cctggcagga 2100 cacaggcact tttccaatgg gcaagcccag tggaggcagg atgggagagc cccctgggtg 2160 ctgctggggc cttggggcag gagtgaagca gaggtgatgg ggctgggctg agccagggag 2220 gaaggaccca gctgcaccta ggagacacct ttgttcttca ggcctgtggg ggaagttccg 2280 ggtgccttta ttttttattc ttttctaagg aaaaaaatga taaaaatctc aaagctgatt 2340 tttcttgtta tagaaaaact aatataaaag cattatccct aaaaaaaaaa 2390 24 1471 DNA Homo sapiens misc_feature Incyte ID No 1597872CB1 24 cttggctagg agcccgctgc gggccaggcg cgcggtgcgc aggcataccc gggtcgcggg 60 tgaacaccgc ggagccccgg gacttgcctg tcgttgctgg tggcggcatg gtcttctgtc 120 tgtcgagcga ggagccgcgc cgcccgctgc gaacgacatg gtccacttcc aggcctcgga 180 agtccagcag ctgctacaca acaagttcgt ggtcatcttg ggggactcca ttcagagggc 240 tgtgtacaag gacctggtgc tcttgctcca gaaagactca ctgctcacag ctgcccagct 300 gaaagccaag taccttgagg atgttctgga agagctgaca tatggacctg ccccggacct 360 ggtgatcatc aactcctgcc tctgggatct ctccagatat ggtcgctgct caatggagag 420 ctaccgggag aacctggagc gggtgtttgt gcgcatggac caagtattgc cagactcctg 480 cctgctggtg tggaacatgg cgatgcccct cggggaacgt atcactgggg gtttcctcct 540 gccagagctc cagcccctgg caggctccct gcggcgggat gtggttgaag ggaacttcta 600 cagtgctacg ctggccgggg accactgctt tgatgtccta gacctccact ttcacttccg 660 gcatgcagta cagcaccgtc atcgggatgg tgtccactgg gaccagcatg cacaccgcca 720 cctctcacac ctgcttctga cccatgtggc tgacgcctgg ggcgtggagc tgcccaagcg 780 tggctatccc cctgacccgt ggattgagga ctgggcagag atgaatcatc cattccaggg 840 aagccatagg cagaccccag acttcgggga gcacctggcc ttgctcccac ccccaccttc 900 ttctttgcct cctcccatgc cttttcccta cccgcttcct cagccctcgc cacctcccct 960 cttcccaccc ctgccccagg ataccccttt tttcccaggc cagcccttcc caccccatga 1020 attcttcaac tataatccag tggaggactt ctcgatgcca ccccacttag gatgtggccc 1080 tggagtgaac tttgtgcctg gccctctgcc acctccaatc cctggcccta atccccatgg 1140 tcagcactgg ggcccagtgg tccaccgggg gatgccacgc tatgttccta acagccccta 1200 ccatgtgcgg agaatggggg ggccctgcag gcagcggctc agacactcag agagactgat 1260 ccacacatac aaactggaca gacggcctcc tgcccattcg gggacatggc ctgggtagac 1320 tggatcttgg gctgggactg gatgtgccaa tggcccttca gggcctgcct ggcacctcag 1380 gtactgggct agggtgtctg ctatgcctgg tattgttctt gtccattgct gtcaccaata 1440 aaggcatgga agaacagagt gaaaaaaaaa a 1471 25 3293 DNA Homo sapiens misc_feature Incyte ID No 1674661CB1 25 gcgagcgcgg gcaggcggcg acgcgggggc aggggtggac ggcggtcaga gccgaacgcg 60 agggcggcgc ccggggactg gagctgcgcg caataggaca gctggcctga agctcagagc 120 cggggcgtgc gccatggccc cacactgggc tgtctggctg ctggcagcaa ggctgtgggg 180 cctgggcatt ggggctgagg tgtggtggaa ccttgtgccg cgtaagacag tgtcttctgg 240 ggagctggcc acggtagtac ggcggttctc ccagaccggc atccaggact tcctgacact 300 gacgctgacg gagcccactg ggcttctgta cgtgggcgcc cgagaggccc tgtttgcctt 360 cagcatggag gccctggagc tgcaaggagc gatctcctgg gaggcccccg tggagaagaa 420 gactgagtgt atccagaaag ggaagaacaa ccagaccgag tgcttcaact tcatccgctt 480 cctgcagccc tacaatgcct cccacctgta cgtctgtggc acctacgcct tccagcccaa 540 gtgcacctac gtcaacatgc tcaccttcac tttggagcat ggagagtttg aagatgggaa 600 gggcaagtgt ccctatgacc cagctaaggg ccatgctggc cttcttgtgg atggtgagct 660 gtactcggcc acactcaaca acttcctggg cacggaaccc attatcctgc gtaacatggg 720 gccccaccac tccatgaaga cagagtacct ggccttttgg ctcaacgaac ctcactttgt 780 aggctctgcc tatgtacctg agactgtggg cagcttcacg ggggacgacg acaaggtcta 840 cttcttcttc agggagcggg cactggagtc cgactgctat gccgagcagg tggtggctcg 900 tgtggcccgt gtctgcaagg gcgatatggg gggcgcacgg accctgcaga ggaagtggac 960 cacgttcctg aaggcgcggc tggcatgctc tgccccgaac tggcagctct acttcaacca 1020 gctgcaggcg atgcacaccc tgcaggacac ctcctggcac aacaccacct tctttggggt 1080 ttttcaagca cagtggggtg acatgtacct gtcggccatc tgtgagtacc agttggaaga 1140 gatccagcgg gtgtttgagg gcccctataa ggagtaccat gaggaagccc agaagtggga 1200 ccgctacact gaccctgtac ccagccctcg gcctggctcg tgcattaaca actggcatcg 1260 gcgccacggc tacaccagct ccctggagct acccgacaac atcctcaact tcgtcaagaa 1320 gcacccgctg atggaggagc aggtggggcc tcggtggagc cgccccctgc tcgtgaagaa 1380 gggcaccaac ttcacccacc tggtggccga ccgggttaca ggacttgatg gagccaccta 1440 tacagtgctg ttcattggca caggagacgg ctggctgctc aaggctgtga gcctggggcc 1500 ctgggttcac ctgattgagg agctgcagct gtttgaccag gagcccatga gaagcctggt 1560 gctatctcag agcaagaagc tgctctttgc cggctcccgc tctcagctgg tgcagctgcc 1620 cgtggccgac tgcatgaagt atcgctcctg tgcagactgt gtcctcgccc gggaccccta 1680 ttgcgcctgg agcgtcaaca ccagccgctg tgtggccgtg ggtggccact ctggatctct 1740 actgatccag catgtgatga cctcggacac ttcaggcatc tgcaacctcc gtggcagtaa 1800 gaaagtcagg cccactccca aaaacatcac ggtggtggcg ggcacagacc tggtgctgcc 1860 ctgccacctc tcctccaact tggcccatgc ccgctggacc tttgggggcc gggacctgcc 1920 tgcggaacag cccgggtcct tcctctacga tgcccggctc caggccctgg ttgtgatggc 1980 tgcccagccc cgccatgccg gggcctacca ctgcttttca gaggagcagg gggcgcggct 2040 ggctgctgaa ggctaccttg tggctgtcgt ggcaggcccg tcggtgacct tggaggcccg 2100 ggcccccctg gaaaacctgg ggctggtgtg gctggcggtg gtggccctgg gggctgtgtg 2160 cctggtgctg ctgctgctgg tgctgtcatt gcgccggcgg ctgcgggaag agctggagaa 2220 aggggccaag gctactgaga ggaccttggt gtaccccctg gagctgccca aggagcccac 2280 cagtcccccc ttccggccct gtcctgaacc agatgagaaa ctttgggatc ctgtcggtta 2340 ctactattca gatggctccc ttaagatagt acctgggcat gcccggtgcc agcccggtgg 2400 ggggccccct tcgccacctc caggcatccc aggccagcct ctgccttctc caactcggct 2460 tcacctgggg ggtgggcgga actcaaatgc caatggttac gtgcgcttac aactaggagg 2520 ggaggaccgg ggagggctcg ggcaccccct gcctgagctc gcggatgaac tgagacgcaa 2580 actgcagcaa cgccagccac tgcccgactc caaccccgag gagtcatcag tatgagggga 2640 acccccaccg cgtcggcggg aagcgtggga ggtgtagctc ctacttttgc acaggcacca 2700 gctacctcag ggacatggca cgggcacctg ctctgtctgg gacagatact gcccagcacc 2760 cacccggcca tgaggacctg ctctgctcag cacgggcact gccacttggt gtggctcacc 2820 agggcaccag cctcgcagaa ggcatcttcc tcctctctgt gaatcacaga cacgcgggac 2880 cccagccgcc aaaacttttc aaggcagaag tttcaagatg tgtgtttgtc tgtatttgca 2940 catgtgtttg tgtgtgtgtg tatgtgtgtg tgcacgcgcg tgcgcgcttg tggcatagcc 3000 ttcctgtttc tgtcaagtct tcccttggcc tgggtcctcc tggtgagtca ttggagctat 3060 gaaggggaag gggtcgtatc actttgtctc tcctaccccc actgccccga gtgtcgggca 3120 gcgatgtaca tatggaggtg gggtggacag ggtgctgtgc cccttcaaaa ggaagtgcaa 3180 ggccttgggg tgggcctagt cttgctcctt agggctgtga atggttttca gaggttgggg 3240 gaaggaaatt ggaaccctcc tgtgtgtttt gggggggaaa gggttggttg ggg 3293 26 1324 DNA Homo sapiens misc_feature Incyte ID No 1689337CB1 26 cagaatatca gatgtccaac atttcccttc tggctctttt tagctcccaa gtatctgctt 60 ccttaaaggc cctgagtcac ttcttctctc tttgcttcag actggcccgg gagcaagcgc 120 gagtgtgcga actgcagagt gggaaccagc agctggagga gcagcgggtg gagctggtgg 180 aaagactgca ggccatgctg caggcccact gggatgaggc caaccagctg ctcagcacca 240 ctctcccgcc gcccaaccct ccagctcctc ctgctggacc ctccagcccc gggcctcagg 300 agcccgagaa ggaggagagg agggtctgga ctatgcctcc catggccgtg gccctgaagc 360 ctgtattgca gcagagccgg gaagcaaggg acgagctacc tggagcgcct cctgttcttt 420 gcagttcctc ctcagatctt agcctcctgt tgggcccctc ttttcagagc cagcattctt 480 tccagcccct ggagcccaaa ccagacctca cttcatccac agctggggcc ttctctgcac 540 ttggggcctt ccatcccgat catagggcag aaaggccatt ccctgaggaa gatcctggac 600 ctgacgggga gggcctccta aagcaagggc tgccgcctgc tcagctggag ggcctcaaga 660 attttttgca ccagttgctg gagacagtgc cccagaacaa tgagaaccct tctgtcgacc 720 tgttgccccc taagtctggt cctctgactg tcccatcttg ggaggaagcc cctcaagtgc 780 cacgtattcc accgcctgtc cacaaaacca aagttccctt agccatggca tccagtcttt 840 tccgggtccc tgagcctccc tcctcccatt cacaaggcag tggtcccagc agtggttccc 900 cagagagagg tggagatggg cttacattcc caaggcagct gatggaggtg tctcaactgt 960 tgcgactcta ccaggctcgg ggctgggggg ctctgcctgc tgaggatctc ctgctctacc 1020 tgaagaggct ggaacacagc ggcaggactg atggccgagg ggataatgtc cccagaagga 1080 acacagactc ccgcttgggt gagatccccc ggaaagagat tccctcccag gctgtccctc 1140 gccgccttgc tacagccccc aagactgaaa aacctcccgc acggaagaaa agtgggcacc 1200 ctgccccgag tagcatgagg agccgggggg gagtctggag atgagccccc ctaccctctc 1260 tcctctttgt tctctcattg ttgttatttt aataaatgct cagtagtctg taaaaaaaaa 1320 aaaa 1324 27 1884 DNA Homo sapiens misc_feature Incyte ID No 1746392CB1 27 cgagggggtg tggaaactta ccggctgagc catggataca ccgttaaggc gcagccgacg 60 gctgggaggc ctaaggcccg aatcccccga gagcctcacc tcagtttcgc ggacgagacg 120 ggcccttgtg gagttcgagt cgaacccaga agaaacgagg gagcccgggt ctcctccgag 180 tgtgcagcgg gctggcctgg ggtcccccga aaggccgccg aagacaagcc caggatcacc 240 ccgtctgcag cagggtgcag gcttggagtc accccaaggg cagccagagc caggcgcagc 300 gtccccccag cgtcagcaag acctacacct ggagtcgcct caaagacagc cagagtacag 360 tcctgaatcc ccacgatgtc agccgaagcc aagtgaggag gcaccgaagt gttctcagga 420 ccagggagta ctggcctcgg agttggccca gaataaggag gagctgaccc cgggggcccc 480 ccagcatcag ctaccgccgg tcccaggatc accagagcct taccccggtc agcaagctcc 540 cggtccggag ccctctcagc cactactgga gctgacaccc agggcacctg gctccccccg 600 gggtcagcat gagccgagca agccacctcc agctggggag acggtgacag gcggcttcgg 660 ggcaaagaag cgaaaaggtt cttcatccca ggccccagcg tccaagaagt tgaataaaga 720 ggagcttcct gtaatcccga aggggaagcc caaatcgggg cgagtgtgga aggaccgctc 780 caagaaaaga ttctcccaga tgcttcagga caagcccctg cgcacatcgt ggcagcggaa 840 gatgaaggaa cgacaggaga ggaagctggc caaggacttt gcccgtcacc tggaggagga 900 gaaggagagg cgccgccagg agaagaaaca gcgccgggct gagaacctga aacgccgcct 960 ggagaatgag cggaaggcag aggtcgtcca agtgatccga aaccccgcca agctcaagcg 1020 ggcaaagaag aagcagctgc gctccattga gaagcgggac accctggccc tgctgcagaa 1080 gcagccgccc cagcagccgg cagccaagat ctgagctcag gacggcccga gggccttcca 1140 tggccaacaa acatgtcaga cacagcacct caggccgctg ctcagatgcc tctgctggag 1200 ctggcactcc aaacccatgg ctccagaaca gggaccccca ccccgaccgg ggctcctcgg 1260 cctttgaagg cttccaggca ggtctgtgtg ggacagaagc ccagaggggg cctgggacct 1320 ggcagagatg ggggcgggaa gagattcagc tcccatccct ccttcctctc cttctccaag 1380 tgccttcaaa ccaagaactg tacattcttc tggttcctca gtgagctggt gactggcagg 1440 tgactccctc agcagtgtat gccctttctc agcatcctag gtccatccca ggcctggagg 1500 ctgacagttg ggaatccagc ttcccccaca ccttcccaaa ggctgctctg agcacctcca 1560 caccccactg cctctgtccc cagcaaactg aatccggttc ctctccactt ttcaatactg 1620 aaagattaaa atggggaggt tgcagggagc agagcttttc cctagcaccc actttcccaa 1680 accagtctct gcagaagccc cagagaatct aactcatgcc tgtccagtct acagcaaaaa 1740 tatttattga gtgcctgttg catacaggca caatcctagg caccggcaaa tacagacaat 1800 agaccaaagt ccctgccctc gaggagcttt cattctgatg gagagaaaac ataataaaca 1860 agcaaaatgc acaaaaaaaa aaaa 1884 28 2429 DNA Homo sapiens misc_feature Incyte ID No 1825182CB1 28 cctatttcca aaaagctcga ctggagtgtt ataaaacctg aaaattctct tgtgctttct 60 cttcttttgc ttctagttac catcctcaaa ggattggcta aaagcaagca actggattga 120 acaccctaag aagaaagatt cacactgcac caggagacat cagaaagaat gaaaactctg 180 ccgctgtttg tgtgcatctg tgcactgagt gcttgcttct cgttcagtga aggtcgagaa 240 agggatcatg aactacgtca cagaaggcat catcaccaat cacccaaatc tcactttgaa 300 ttaccacatt atcctggact gctagctcac cagaagccgt tcattagaaa gtcctataaa 360 tgtctgcaca aacgctgtag gcctaagctt ccaccttcac ctaataagcc ccccaaattc 420 ccaaatcctc accagccacc taaacatcca gataaaaata gcagtgtggt caaccctacc 480 ttagtggcta caacccaaat tccatctgtg actttcccat cagcttccac caaaattact 540 acccttccaa atgtgacttt tcttccccag aatgccacca ccatatcttc aagagaaaat 600 gttaacacaa gctcttctgt agctacatta gcaccagtga attccccagc tccacaagac 660 accacagctg ccccacccac accttctgca actacaccag ctccaccatc ttcctcagct 720 ccaccagaga ccacagctgc cccacccaca ccttctgcaa ctacacaagc tccaccatct 780 tcctcagctc caccagagac cacagctgcc ccacccacac ctcctgcaac tacaccagct 840 ccaccatctt cctcagctcc accagagacc acagctgccc cacccacacc ttctgcaact 900 acaccagctc cactatcttc ctcagctcca ccagagacca cagctgtccc acccacacct 960 tctgcaacta ccctagaccc atcatccgcc tcagctccac cagagaccac agctgcccca 1020 cccacacctt ctgcaactac accagctcca ccgtcttccc cagctccaca agagaccaca 1080 gctgccccaa ttaccacacc taattcttcc ccaactactc ttgcacctga cacttctgaa 1140 acttcagctg cacccacaca ccagactact acttcggtca ctactcaaac tactactact 1200 aaacaaccaa cttcagctcc tggccaaaat aaaatttctc gatttctttt atatatgaag 1260 aatctactaa acagaattat tgacgacatg gtggagcaat agtatattgt atgttgtaaa 1320 gtgttctgtc atttacaaga tgtgattcat gagtgcagaa ctaccacctt tcttttagca 1380 ccaatcccaa catgaaatta tattactcag atttaaagca ctatcattaa tctttcaatc 1440 taattattca ccaccacaag acctattaac aagacaaaat gcctctatcc cacaagccag 1500 atgcaggtct ggggttcaaa ataactcttt ggatcctaca gagatagcct actgagggca 1560 gagaaagtcc ttagataaag agagaatatt gtatgggcca tcaaccattt acttttccct 1620 gaatgttaga aactacaaaa ccactacctt gtacccccat caaaatccca cctgaaccat 1680 ctaatcctat aaacataaag gggtaaaatt ggaactctcc agatgaacaa agacatctaa 1740 atatctgtag atagaaacat ttatctatct aaatatattg atagacctgt cattgtattg 1800 attaatgaca aaacccttta gataattatc ttccatttta aataaaattt tatttcacaa 1860 atatgagcca agaaagagga aagttgattt gaagtgagga ttagaagtga atgacaataa 1920 agtctggcag ccaagcacga accaagactg gcactatttt tcttagtgta tataattgtt 1980 taaactgcaa ggttgacatt tattgtgttg tgtctaagtt aattttgatc taatgtacct 2040 gattctagcc tctgtgaaca acaagaatat gtttgtgtat gttcacatgg tgcttataat 2100 atttcactat caattcaatt aattcacata aattccatgt gaaatgtatt caacaatgga 2160 atattttcta aaacatttag tatacatttg aatgtatttt aaaccatgcc aaactactgc 2220 tttaatgtca agtttgcaga attgtctctg aaaataaaaa ccctgacttt agttgtaaaa 2280 caataaaagt tagctacttg gtatacggag atgttaattt gggatatgga ggcattttta 2340 tcttctgtca ctactactta aaactctgat gattatgtta gatttttttg ctaactaata 2400 aagatttcaa atggcaattt ataaaaaaa 2429 29 985 DNA Homo sapiens misc_feature Incyte ID No 2155541CB1 29 ggaagagggc ggcgacggtg gtggtgactg agcggagccc ggtgacagga tgttggtgtt 60 ggtattagga gatctgcaca tcccacaccg gtgcaacagt ttgccagcta aattcaaaaa 120 actcctggtg ccaggaaaaa ttcagcacat tctctgcaca ggaaaccttt gcaccaaaga 180 gagttatgac tatctcaaga ctctggctgg tgatgttcat attgtgagag gagacttcga 240 tgagaatctg aattatccag aacagaaagt tgtgactgtt ggacagttca aaattggtct 300 gatccatgga catcaagtta ttccatgggg agatatggcc agcttagccc tgttgcagag 360 gcaatttgat gtggacattc ttatctcggg acacacacac aaatttgaag catttgagca 420 tgaaaataaa ttctacatta atccaggttc tgccactggg gcatataatg ccttggaaac 480 aaacattatt ccatcatttg tgttgatgga tatccaggct tctacagtgg tcacctatgt 540 gtatcagcta attggagatg atgtgaaagt agaacgaatc gaatacaaaa aaccttaaag 600 ccaggcctgt cttgatgatt tttggttttt tttcattgtc ctgttgaaat caagtaatta 660 aacatttaag agccacaaaa ttgtatcact tttataatat tttgcagtaa aatataatac 720 catcttctct gttaatacat aattgctcca agcttcctgt aaactataag aatatattta 780 gtttacagta tatggattct atgaaaaaat gtccacaaca cagtaattgg tcacttgtta 840 agaaaaattt atccttgtaa gtatcttcaa agttgatatt tggaacttta ttccaaaagt 900 agtgcatgtg gagaaagaat ctagactttc ttgtatacat ttttctcttc tccagtaata 960 aacaattacc tttcatttaa aaaaa 985 30 3381 DNA Homo sapiens misc_feature Incyte ID No 2215706CB1 30 gttagctcag tgtaacattg atgagctgaa gaaagagatg aatatgaatt ttggagactg 60 gcaccttttc agaagcacag tactagaaat gagaaacgca gaaagccacg tggtccctga 120 agacccacgt ttcctcagtg agagcagcag tggcccagcc ccgcacggtg agcctgctcg 180 ccgcgcttcc cacaacgagc tgcctcacac cgagctctcc agccagacgc cctacacact 240 caacttcagc ttcgaagagc tgaacacgct tggcctggat gaaggtgccc ctcgtcacag 300 taatctaagt tggcagtcac aaactcgcag aaccccaagt ctttcgagtc tcaattccca 360 ggattccagt attgaaattt caaagcttac tgataaggtg caggccgagt atagagatgc 420 ctatagagaa tacattgctc agatgtccca gttagaaggg ggccccgggt ctacaaccat 480 tagtggcaga tcttctccac atagcacata ttacatgggt cagagttcat cagggggctc 540 tattcattca aacctagagc aagaaaaggg gaaggatagt gaaccaaagc ccgatgatgg 600 gaggaagtcc tttctaatga agaggggaga tgttatcgat tattcatcat caggggtttc 660 caccaacgat gcttcccccc tggatcctat cactgaagaa gatgaaaaat cagatcagtc 720 aggcagtaag cttctcccag gcaagaaatc ttccgaaagg tcaagcctct tccagacaga 780 tttgaagctt aagggaagtg ggctgcgcta tcaaaaactc ccaagtgacg aggatgaatc 840 tggcacagaa gaatcagata acactccact gctcaaagat gacaaagaca gaaaagccga 900 agggaaagta gagagagtgc cgaagtctcc agaacacagt gctgagccga tcagaacctt 960 cattaaagcc aaagagtatt tatcggatgc gctccttgac aaaaaggatt catcggattc 1020 aggagtgaga tccagtgaaa gttctcccaa tcactctctg cacaatgaag tggcggatga 1080 ctcccatctt gaaaaggcaa atctcataga gctggaagat gacagtcaca gcggaaagcg 1140 gggaatccca catagcctga gtggcctgca agatccaatt atagctcgga tgtccatttg 1200 ttcagaagac aagaaaagcc cttccgaatg cagcttgata gccagcagcc ctgaagaaaa 1260 ctggcctgca tgccagaaag cctacaacct gaaccgaact cccagcaccg tgactctgaa 1320 caacaatagt gctccagcca acagagccaa tcaaaatttc gatgagatgg agggaattag 1380 ggagacttct caagtcattt tgaggcctag ttccagtccc aacccaacca ctattcagaa 1440 tgagaatcta aaaagcatga cacataagcg aagccaacgt tcaagttaca caaggctctc 1500 caaagatcct ccggagctcc atgcagcagc ctcttctgag agcacaggct ttggagaaga 1560 aagagaaagc attctttgag aaaaacaagc aaaggagaag agtgttactg tacccttatg 1620 acagaattgt cctggatttt gactccatcc acgcccatca cctttctaca ttttgctgac 1680 agataactaa ccgatgatga ggccgaggta aaagagacat ctgcagtgtg acagaaggga 1740 gcatgagaag catggctcac cagccagcct ctgtggtctt tgtaattaga agcttcagaa 1800 ctcactaata ctactgtacc tttcattggc gcattacccc ataaaacttt ttgagacgag 1860 gtgagatctg agtataaaga taggtcagaa gtattttaaa gggcttaatg tgccaaaaag 1920 aaaaaaagct agagaccctt tttgcaaaca tttggtgacc acacatttga gggaagacgt 1980 ggcgttaggt gaagcagaag caaaccctgc tcttaggggc tcacctaggt gagtgcacag 2040 cctgtgacgc tacagggaga ggctgagtaa accgagatcc agcgttctgt atggcagggg 2100 tattgcttat cacagaggtt ctgaagagta ggaagtacat aatgaagagg gctttaaaaa 2160 ttgccaacaa agtgagtcac cagggctggc agtagtgtga cggggctgtc ctgagctgtt 2220 aggagagtag atgcggggag ggctggtgac ctccgtgggt ttatatgtcg gaaactcttc 2280 tctccaaatc ccaggcctgg cttccagcac catccagctg tgcccaagaa gccaccctgg 2340 tctgttctcc aactctttta aatggtgccc aacttttcta agtgagctta gcaatgagaa 2400 gaaaaaaaaa catgaattct ttttctggaa aatcagggag acatgggtaa taataggtac 2460 taataaatat ttatagatga gtgaatgagg aaataattac atcaaaaagg tcagtgacaa 2520 ttgataaatg acaaggaaat atttaattag gtaaaactaa atcattgctc tctatactag 2580 gatagacttt atctacttca tctgttccta agtcagcatg ttagttctgg ggaaggatca 2640 taagaaagga aatacttttt aaaaaaaaat ttggaaacat gtaacaaagc aagggtaaaa 2700 tatatatata tatctatata agtgctgtga ctgtaaaagt gtactttcca ttaattatta 2760 gccgagttaa gagaatggtc acattgaagt actgtgtgga ctagaaatgt accctgtcat 2820 catgcaatga aatattgtta tcgttttaac atagctcatt tatgtagaat gaattctggt 2880 ggtttacccc aagtcacagt taggacggta gatggtgaga tcgcagatgc gctattatct 2940 agattcagtg ttacattttc gatgtttatc actcagtggg tttttattaa tatgctgatt 3000 aagttattta ctgggccagt cattgtgcta aatagttgct cttttgtgtt tcattgcctt 3060 gatgtttgag tgtaatctag cattttaata cagtgtttat tttgcatgat ctttaacaaa 3120 tgttttaagc aattttaaaa aggcaggatg ttattgacat tatacactga agtcttaaca 3180 ttttaacatt tatagtgctt atttgcaaaa ttgtataatt aggaattatt tcagagacaa 3240 tgttttcttt ttcaggtgag tagttgccgc gtaatatcat tggagtacat tctttatact 3300 gtttgtgaaa ttaatactag catattaagt gtacaaatag atttagaaaa caataaaaaa 3360 ttgcatgcta aaaaaaaaaa a 3381 31 1803 DNA Homo sapiens misc_feature Incyte ID No 2347692CB1 31 gggctgttcc cggggaggct gtgatgggtt gacaggtgcg tgacagtggg agctgctctc 60 ggcacaagca tgtacggcaa aggcaagagt aacagcagcg ccgtcccgtc cgacagccag 120 gcccgggaga agttagcact ctacgtatat gaatatctgc tccatgtagg agctcagaaa 180 tcagctcaaa catttttatc agagataaga tgggaaaaaa acatcacatt gggggaacca 240 ccaggattct tacattcttg gtggtgtgta ttttgggatc tctactgtgc agctccagag 300 agacgtgaaa catgtgaaca ctcaagtgaa gcaaaagcct tccatgatta cagtgctgca 360 gcagctccca gtccagtgct aggaaacatt cccccaggag atggcatgcc agtaggtcct 420 gtaccaccag ggttctttca gccttttatg tcacctcggt accctggagg tccaaggccc 480 ccattgagga tacctaatca ggcacttgga ggtgtcccag gaagtcagcc attactcccc 540 agtggaatgg atccaactcg acaacaagga catccaaata tgggtgggcc aatgcagaga 600 atgactcctc caagaggaat ggtgccctta ggaccacaga actatggagg tgcaatgaga 660 cccccactga atgctttagg tggccctgga atgcctggaa tgaacatggg tccaggtggt 720 ggtagacctt ggccaaaccc aacaaatgcc aattcaatac catactcctc agcatctcct 780 gggaattatg taggtcctcc aggaggtgga gggccaccag gaacacccat catgcctagt 840 ccagcagatt caaccaactc tggtgataac atgtatactt taatgaatgc agtacctcct 900 ggacctaaca gacctaattt tccaatgggt cctgggtcag atggtcccat gggtggatta 960 ggaggaatgg agtcacatca catgaatggc tctttaggct caggagatat ggacagtatt 1020 tccaagaatt ctcccaataa tatgagcctg agtaatcaac cgggcactcc aagggatgat 1080 ggcgaaatgg ggggaaattt cttaaatcct tttcagagtg agagttactc ccctagcatg 1140 acaatgagcg tgtgatccat taccaagtct cctcatgaaa accacagtga gtcagccctt 1200 cacagaacta ctacggaaga aaattattca tcacagtgta cagttaaaca aaggaatctc 1260 agtcacacca aaccaacctt tttatttcct gctctctccc ctcttttgtg aagacagcgg 1320 gtccaaatgt gattcaaaca actgtacgga gtggcatatt agaattgccc taaactgaac 1380 tgcaaataat tatgtgtgta tgtatatgtg tgggaaagag aatgtactgt atatgtgtat 1440 gttatacaga catatacaca tacatacatt gacccacagg acattgtaaa atattatcac 1500 atgacatctt aagtagaaat aagtagggac ttttattcca tccttttttt cacgtttaca 1560 ttttaattat tacaagttgc tcctgccccc tccctgaact attttgtgct gtgtatatca 1620 ctgctttata taagttattt tttaaggtga actcagatgt tatggttttg taaatgtctg 1680 caatcatgga taggaataaa atcgcttatt tgagagcttt cattaaattg cgtctgatgc 1740 aagttatcct gtgaatcaca aagtgtactg tctcaaaaaa tagaagaaaa tgtgaaaaaa 1800 aaa 1803 32 1515 DNA Homo sapiens misc_feature Incyte ID No 2579048CB1 32 ggtaaagtgg cttctgggcg gaaggtacac tataggctcg gggaggtaag cggcggcagg 60 ccggcggttg gtgtgtcccg ggtgtgggga ggcgacagag ccctggcact tgagggttga 120 gggggcctcc ccagcgcggc gaaccgtcta gcctccggag gccaggccgt gagtgcggga 180 ggtatacgcc aaggcggaag aattttgcca ctcactacct gtgtgacctc gggtaaatta 240 gccttggaac gtcagtttct tcgtctctat aattgaaata ataatagtac ctctctcagg 300 attgttgtga gccgtcagtg aaacacttag agcagtttct ggcacatggt agaattgggc 360 tatttgctga agcttcttgg tggcccttgc tagcccagga agaaacttac attttgattt 420 ttttgtacca tggctttggt tcacaaattg ctgcatggta cttattttct cagaaaattc 480 tctaagccaa cttctgcctt gtatccattt ttgggtattc tctttgcaga gtattccagt 540 agtcttcaga aaccagtggc ttctcctggc aaagcctcct cacagaggaa gactgaaggg 600 gatttgcaag gagatcacca gaaagaagtt gctttggata taacttcttc tgaggagaag 660 cctgatgtta gtttcgataa agcaattaga gatgaagcaa tataccattt taggcttttg 720 aaggatgaaa ttgtggatca ttggagagga ccggaaggcc accctctgca tgaggtcttg 780 ctggaacaag ccaaggttgt ctggcaattc cgggggaaag aagatttgga taagtggaca 840 gtgacttctg ataagacgat tggaggcaga agtgaagtgt ttttgaaaat gggcaagaat 900 aaccaaagtg cactgctata tggaactctg agctctgagg cgcctcagga cggggagtct 960 acccgaagtg ggtactgtgc aatgatatcc aggattccaa ggggtgcttt tgagaggaag 1020 atgtcttacg attggtccca gttcaatact ctgtatctcc gtgtacgtgg ggatggtcgg 1080 ccttggatgg tgaatatcaa ggaggacaca gatttcttcc agaggacgaa tcagatgtat 1140 agttacttca tgttcacccg cgggggaccc tactggcagg aggtcaagat tcctttttcc 1200 aaatttttct tctctaatcg aggaagaatc cgggatgttc agcatgagct tccgcttgat 1260 aagatctctt ctataggatt caccttggct gataaagtgg atggtccatt cttcctggag 1320 atagatttta ttggcgtgtt tactgatcca gctcatacag aagaatttgc ctatgaaaat 1380 tctccagagc ttaacccaag gctttttaaa taaagatcat atggtagttt tgttttacta 1440 atctaagggt actagcatct acaatgatat agacaaaata aaatatttct ttaatggcat 1500 ccaaaaaaaa aaaaa 1515 33 4416 DNA Homo sapiens misc_feature Incyte ID No 2604493CB1 33 cggatgggcg ggcagccgcg ccgccgcggc acttttttaa ttttttcggg tgccgcagcg 60 gcgacccctc ggcgccgatg tccctgatcc ctggagcgac gacggccgct gcctaagctg 120 gaaagaggaa tgccagctcc tgagcaggcc tcattggtgg aggaggggca accacagacc 180 cgccaggaag ctgcctccac tggcccaggc atggaacccg agaccacagc caccactatt 240 ctagcatccg tgaaggagca ggagcttcag tttcagcgac tcacccgaga actggaagtg 300 gaaaggcaga ttgttgccag tcagctagaa agatgtaggc ttggagcaga atcaccaagc 360 atcgccagca ccagctcaac tgagaagtca tttccttgga gatcaacaga cgtgccaaat 420 actggtgtaa gcaaacctag agtttctgac gctgtccagc ccaacaacta tctcatcagg 480 acagagccag aacaaggaac cctctattca ccagaacaga catctctcca tgaaagtgag 540 ggatcattgg gtaactcaag aagttcaaca caaatgaatt cttattccga cagtggatac 600 caggaagcag ggagtttcca caacagccag aacgtgagca aggcagacaa cagacagcag 660 cattcattca taggatcaac taacaaccat gtggtgagga attcaagagc tgaaggacaa 720 acactggttc agccatcagt agccaatcgg gccatgagaa gagttagttc agttccatct 780 agagcacagt ctccttctta tgttatcagc acaggcgtgt ctccttcaag ggggtctctg 840 agaacttctc tgggtagtgg atttggctct ccgtcagtga ccgacccccg acctctgaac 900 cccagtgcat attcctccac cacattacct gctgcacggg cagcctctcc gtactcacag 960 agacccgcct ccccaacagc tatacggcgg attgggtcag tcacctcccg gcagacctcc 1020 aatcccaacg gaccaacccc tcaataccaa accaccgcca gagtggggtc cccactgacc 1080 ctgacggatg cacagactcg agtagcttcc ccatcccaag gccaggtggg gtcgtcgtcc 1140 cccaaacgct cagggatgac cgccgtacca cagcatctgg gaccttcact gcaaaggact 1200 gttcatgaca tggagcaatt cggacagcag cagtatgaca tttatgagag gatggttcca 1260 cccaggccag acagcctgac aggcttacgg agttcctatg ctagtcagca tagtcagctt 1320 gggcaagacc ttcgttctgc cgtgtctccc gacttgcaca ttactcctat atatgagggg 1380 aggacctatt acagcccagt gtaccgcagc ccaaaccatg gaactgtgga gctccaagga 1440 tcgcagacgg cgttgtatcg cacaggttca ggtattggaa atctacaaag gacatccagc 1500 caacgaagta cccttacata ccaaagaaat aattatgctc tgaacacaac agctacctac 1560 gcggagccct acaggcctat acaataccga gtgcaagagt gcaattataa caggcttcag 1620 catgcagtgc cggctgatga tggcaccaca agatccccat caatagacag cattcagaag 1680 gaccccaggg agtttgcctg gcgtgatcct gagttgcctg aggtcattca catgcttcag 1740 caccagttcc catctgttca ggcaaatgca gcggcctacc tgcagcacct gtgctttggt 1800 gacaacaaag tgaagatgga ggtgtgtagg ttagggggaa tcaagcatct ggttgacctt 1860 ctggaccaca gagttttgga agttcagaag aatgcttgtg gtgcccttcg aaacctcgtt 1920 tttggcaagt ctacagatga aaataaaata gcaatgaaga atgttggtgg gatacctgcc 1980 ttgttgcgac tgttgagaaa atctattgat gcagaagtaa gggagcttgt tacaggagtt 2040 ctttggaatt tatcctcatg tgatgctgta aaaatgacaa tcattcgaga tgctctctca 2100 accttaacaa acactgtgat tgttccacat tctggatgga ataactcttc ttttgatgat 2160 gatcataaaa ttaaatttca gacttcacta gttctgcgta acacgacagg ttgcctaagg 2220 aacctcagct ccgcggggga agaagctcgg aagcaaatgc ggtcctgcga ggggctggta 2280 gactcactgt tgtatgtgat ccacacgtgt gtgaacacat ccgattacga cagcaagacg 2340 gtggagaact gcgtgtgcac cctgaggaac ctgtcctatc ggctggagct ggaggtgccc 2400 caggcccggt tactgggact gaacgaattg gatgacttac taggaaaaga gtctcccagc 2460 aaagactctg agccaagttg ctgggggaag aagaagaaaa agaaaaagag gactccgcaa 2520 gaagatcaat gggatggagt tggtcctatc ccaggactgt cgaagtcccc caaaggggtt 2580 gagatgctgt ggcacccatc ggtggtaaaa ccatatctga ctcttctagc agaaagttcc 2640 aacccagcca ccttggaagg ctctgcaggg tctctccaga acctctctgc tggcaactgg 2700 aagtttgcag catatatccg ggcggccgtc cgaaaagaaa aggggctccc catccttgtg 2760 gagcttctga gaatggataa cgatagagtt gtttcttccg tggcaacagc cttgaggaat 2820 atggcactag atgttcgcaa caaggagctc ataggcaaat acgccatgcg agacctggtc 2880 aaccggctcc ccggcggcaa tggccccagt gtcttgtctg atgagaccat ggcagccatc 2940 tgctgtgctc tgcacgaggt caccagcaaa aacatggaga acgcaaaagc cctggccgac 3000 tcaggaggca tagagaagct ggtgaacata accaaaggca ggggcgacag atcatctctg 3060 aaagtggtga aggcagcagc ccaggtcttg aatacattat ggcaatatcg ggacctccgg 3120 agcatttata aaaaggatgg gtggaatcag aaccatttta ttacacctgt gtcgacattg 3180 gagcgagacc gattcaaatc acatccttcc ttgtctacca ccaaccaaca gatgtcaccc 3240 atcattcagt cagtcggcag cacctcttcc tcaccagcac tgttaggaat cagagaccct 3300 cgctctgaat acgataggac ccagccacct atgcagtatt acaatagcca aggggatgcc 3360 acacataaag gcctgtaccc tggtaagacg ccagttgggt gtgtgataca gtctcttgaa 3420 aagccgcatt tccaggcgct tggccagtgg cctgggaagt agcctgtgct tgtattgaga 3480 cagtccccca gcagcaaacc atgttccagt cattcccttt cctactttgg ggattgttgc 3540 cttttctgct tgtttaaagt aaaacaagca tgtacttgtt tgtatgtatg tatgtatgta 3600 gttgtacggt gggcacaaat aaaaagaggg ctgtatccaa ataaatcatt tctggctgct 3660 cactggcaca gtccctttgc tccgtcccct cctggctcga gcagtctctg tgttttccac 3720 atgcatcaga accgtcagcc cagtgtcact ttgcaggggc cacatcttct agactggctc 3780 atctcttaaa ttcaaaccag agaatggaga tcaatttcat ttacattttc atggagaaga 3840 gtttgaataa atagcaaaag gtataatgtg attcttctca acgtgattat tttttaagaa 3900 taacaaacaa aaataatgtt acatagtgtt aatattttta tatcctattc agaaagctgt 3960 gcatacaatt tatttaaagg ataccagccc aaagggttac taagatttca tactgttccc 4020 attctcaacc gtctggcgat tataactact ttagatgttc caaataagtc agtgtgagct 4080 accccacaaa aagtactctt ttctcaaacc tgtctattaa aaataaataa aagctggtgg 4140 cagctttact cctgctgggg gggcagtctc cattgtactg ttgtctttga aattgtgatt 4200 tcacaggact atttactaat cgactgttgt tactatggca attaatagct cagaaaattg 4260 cagatgctac atttcaacac caagctctgg gcaaaaggaa atgcacatac caactgaagt 4320 gcatggttgc ttccgatttg cagcagcctg ctttatctta cagatcacct ctttagcatg 4380 catgcccttc tgtatcttct tcgcttgctt caataa 4416 34 4428 DNA Homo sapiens misc_feature Incyte ID No 2787182CB1 34 cggagggccg gggctgggtg gcagcggcca tggaccgctt cgtttggacc agcggcctcc 60 tggagatcaa cgagaccctg gtgatccagc agcgcggggt gcgaatctac gatggcgagg 120 agaagataaa atttgatgct gggactctcc ttcttagtac acaccgactg atttggagag 180 atcagaaaaa tcatgagtgt tgcatggcca ttctcctttc ccaaattgtg ttcattgaag 240 aacaggcggc tggaattggg aagagtgcca aaatagtggt tcatcttcac ccagctcctc 300 ctaacaaaga acctggccca ttccagagta gtaagaactc ctacatcaaa ctctccttca 360 aagaacatgg ccagattgag ttttacaggc gtttatcaga ggaaatgaca caaagaagat 420 gggagaatat gccagtttcc cagtcattac aaacaaatag aggaccccag ccaggaagaa 480 taagggctgt aggaattgta ggtattgaaa ggaaactgga agaaaaaaaa aaagaaactg 540 acaaaaacat ttctgaggcc tttgaagacc tcagcaaact aatgatcaag gctaaggaaa 600 tggtggaatt atcaaaatca attgctaata aaattaaaga caaacaaggt gacatcacag 660 aagatgagac catcaggttt aaatcctact tgctgagcat gggaatagct aacccagtta 720 ccagagaaac ctacggctca ggcacacagt accacatgca gctggccaaa caactggctg 780 gaatattgca ggtgccttta gaggaacgag ggggaataat gtcactcacg gaggtgtact 840 gcttagtaaa ccgagctcga ggaatggaat tgctctcacc agaagattta gtgaatgcgt 900 gcaagatgct ggaagcactg aaattacctc tcaggctccg tgtgtttgac agtggcgtca 960 tggtaattga gcttcagtct cacaaggaag aggaaatggt ggcctcggcc ctggagacag 1020 tttcagaaaa gggatcccta acatcagaag agtttgctaa gcttgtggga atgtctgtcc 1080 tcctagccaa agaaaggttg ctgcttgcag agaagatggg ccatctttgc cgtgatgact 1140 cagtggaagg cctgcgtttt tacccaaatt tatttatgac acagagctaa gggttttgta 1200 tttaaaatcc tttttgtcca tatgcttgcg tcatgtagag gttgtatgac attgagctaa 1260 gagataaacc ccgatcaatt gagaatttat tagaacttca cagtgcaatg taaatctctt 1320 ttaatttctc actaaatatg gtccaggaaa tttatttagt atacgcatag gaaaattcag 1380 aaaagtgaat gccaatatga atttaaaatc atgctatagt gcagaaccct cagagtttaa 1440 cttggaatat agtggatttt aacttgatcc tcaaatctaa tcattttata aagaagggaa 1500 tttagttttg cagagaataa aaagagaagt tgcatgttca gacaggttag attattattt 1560 tggtgtaact gaaattcact gattgcacat gacaatgttg ggacaaaata tactgcagca 1620 tgctatatga ggctcctccc cagggctttt agaagcagtc atagacatgt cttcaacata 1680 ccaaataaaa tacctttaaa aatgaaataa ttttatttga cacatatatt tatatatatt 1740 ctatctagtt tctctttgtt tttttaagtg atgatttcat gactggcatt taaagaatgc 1800 aactgtgtca ttttgtttcc aaatgctgtg gattttgaaa ctgaactaga gagctgtata 1860 gacatgccca gagttatgat tacaaattta ggaggtagac ggctcaggaa ttccctggga 1920 ttgttgtgct ggtggaatgg cagagggaac ttcacaggaa ccttagtgtt cttttacctc 1980 aaagccacag acaggaaata gaaagtggaa aagtaatatc tccttttctt ttccataagg 2040 agtttcaaca ctgaacttta aaaagtctat catattccag caatattttt tctttgtcct 2100 ttatgttgta agttgtgtgg aaaaactact tcggtaagaa atgttactga gataacaaca 2160 actggctaat actgcatgta gattgcttag gttttaaagt gactgcctga cttcacatgt 2220 tattgctaca gcctccagta tgttcgcatt atctcaaact cagggacccc acaggacagg 2280 agacaccctt tctgaaactg agttggaagt gaaagggtgg tgatggtttt ggccaagcct 2340 gcgggaggga aagtattgta ttgggagcac ccttgggacc aggaagaggg atgcccaggt 2400 tcacactctg ggacccctaa gacattgtca gtgggtaagg tggagggcca ctgccagagg 2460 ctgctgcagt gccctctggg aagtgccagg gaccctcagc tgcagaccca ggggatccca 2520 atcccttatc tctgcctgaa aagtggctat tgctgcttcg tatcctccag tgtcacagga 2580 atgctgtgcc tgagacatcc cactttagtt tttgttttcc aaacctgtca ctgactcttc 2640 attgtggtga cagatcctct ggttccccca ttgctcagcc tcaggtttat tttgagggta 2700 actcagtgtc tgactgccca agtcttttag gggttagttg gaccattgtg taagcttgtt 2760 tgtatgtctc acctctttct ggttggtact tactgtctca cactgctctg gcaaacttgt 2820 acacacacac acactctctc tctctctcac ctggctaagg cttttaaaat tatgaagata 2880 aataatctgt ttcaccagct ggagtgagtt gcaaggaaga ttgctggagc tactcaatct 2940 agcgctaatg gtttggattc attactgcaa acctacataa tttaacatat ttgtttactt 3000 tagtgtgaca actgatgaaa aaaaatggag caatctgaat tgtataaaat aacttaagaa 3060 ggaagaaaag tgatatataa atatattttg caaatgtcac attaatttaa aaatgagtat 3120 gattgatttt atttttaaag tgggcattct tcactgtttc gagacctttg tatgtatttg 3180 tgtattttta tctttttttt tcaggccatt attataaggt gttattttgg ccctctaatg 3240 tagaagttat gtttaaatac aaccaatcag gccctaagtg caaaaaagtt ctgtcttggt 3300 gcttatcact ggtataatta ttattttttt gttttcctaa ctttgctctt aggagcatga 3360 gcctctttgt agctttatgg taatgcaata tttctggtca tttactgtca aaaaattttt 3420 ttacattgtg ttagtgaaga ctgtgttttc agggttaaat gattggtatc aatgtatata 3480 gagtaaaatt attgcaaaat ttaaaaatga tttttcttgg cattcttttt aaatactctg 3540 aaacatatat gcaaatgaga aacctttaat catactaaag ccagttatgt taagaatttt 3600 tcctttggat tttataatta agagttgctt ataaatactt atgacattga gctcttactg 3660 ttttagttgt cttaaaagaa tgggacatgt catgaactgg tttctttttt taaaaaattc 3720 ttgatgtgaa ttccactgtt acctgaccat gtatattcag atgaatgtac tctttcttgt 3780 tccttgttac cagattaatc tgagagaatg tggattcatc ctgtcatctt tctcatactc 3840 aaatacttaa aatggttctt ttgcccaaaa gtctcagagt cttcattttg gtacagtgga 3900 gtcacaactt gtaaggtggt taggttttaa agaccgtgcc taaggacaaa attgagcatg 3960 ctcagaatgg tccttgaaaa tctcctaaat taccaataaa tttctctttg tagtcttaaa 4020 gtttaaatcc ctgtggagtg atatgtaggt atgaatatat tatgcttata ttaagtacaa 4080 aaatacaaac tgcatatcaa gagattctta tagcattaat aatttccatg catgtgtctt 4140 tttccagtag gtatggttga atttatgtaa atttattgct aatcccatcc cttacgatct 4200 agagtataag ctgcgcaagg gcagaagttt ttatctggtt tgttcatgga tgtattctaa 4260 gagctgagaa cagggcctgg acacaataag cattcaataa atatttactg aatgaatgaa 4320 ctcctaccta tattcctatt tataatttgg ctccacttta tcctacttta gctcccattc 4380 aattcaataa aaaaaacatt tttttgaaca cataaaaaaa aaaaaaaa 4428 35 1907 DNA Homo sapiens misc_feature Incyte ID No 3096668CB1 35 gcgttgctgc gtaaatggcg ggggcgtgtc ttttggctcc tccgcgtgta gttacctgag 60 aaacgcggga agttgggccc aggcagtgtt gctgcggttg cctaagttgt tttactattt 120 ctggagagag ccgtgagctt gtccaggggc cccaatcctg aggccgaccc ggtttctggc 180 gcggtgcgat ggaggtagtg gaggccgccg ccgctcagct ggaaactctg aaattcaatg 240 gcaccgactt tggagttggg gaaggtccgg cggctccgtc tccgggctct gcccctgtgc 300 cagggacaca gccgccgcta cagtcgtttg aggggtcccc ggacgctggg cagaccgtgg 360 aggttaagcc tgccggggag cagcctctgc agcccgttct gaacgccgtc gcggccggga 420 ccccggcgcc gcagccacag ccaccggctg aatcgccggc ctgcggagac tgcgtcacct 480 ccccaggagc cgcagagcct gcgcgggcgc cggactcctt ggagacctcg gactcggatt 540 cggactcgga cagtgaaaca gattcagata gttcaagttc atcgtcttcc tcttcatctt 600 cctcatcgtc gtcttcttcc tcttgtatat cacttcctcc agtgctgtca gatggagatg 660 atgatttaca agttgagaag gaaaataaga attttcctct taaaacaaaa gatgaattac 720 ttcttaatct atagagaaaa aaaaaattca gtcatggctc tcagtactcc agggattgga 780 tcctactttc caaccatatc tgaaaagtga aggctcacct aatagcatca ggtaattggc 840 tgccacgtgc atgtcctgtg ttgggtaata tgagaaaagt ggttagataa tttcatccca 900 ccttcctcac ttttctcaat tctccccaaa ctgacaccaa aaaggtttag ttttcatttg 960 gaatgtttga ttccttctcc caccaacctc tgttgtcctt tttccatatt tgtgagtgaa 1020 aatcctactg aaagttcttt atataatttt actttagaaa gtcatcaatt tactgatact 1080 cttttttgct caataagcct tcacaaatgc ttgaaaagtc aacagtgtgg taaatgataa 1140 atgttacctt ttcagagggc cacttttgac atattagctt cttccatttt tgcattgaaa 1200 gtgttaggca cccatttttt gactggtatg tctataacct catttttaat taggatggat 1260 tttttaatgt tatgtggtac tcttttgaat attttgctta actgaataac cttgtagacc 1320 atacagatgt tagaaaatta aaatataata atcttacaaa aacagacaat aattgctctt 1380 tcatggatat tccagaagtg tggttaggat cttgcatttc tttaatgcca tcgttacgag 1440 tatctgttat cacagaagca cagtaatttt tttattggga tctagtatat aatttctgta 1500 ttaatgtgtt gtctttttga ttacatgcag ttggacattt aaattgtcgt tagccatttg 1560 tgtaactgtt tagaacttgt gctcagtttc cttactattc gtaggtaaga aaaatgaaac 1620 cagaaagaaa gagacttgtc tgaagtcata ttagtgggga ggagcagatc tgaactggta 1680 atagatacat tccaaactgc tgtgttttta gtatttgtgg agacctgcat agcaacgttg 1740 gaatctggtc gtttggactt tctcattatt tggtttctga ataatcatgt agcatatttt 1800 ctcaaagtga aggatgaatg tatcatctaa tgtaagcttt ttaaaattgt atttgcttgg 1860 attatattgc ctctcaataa agcttttgac ttggaaaaaa aaaaaaa 1907 36 1839 DNA Homo sapiens misc_feature Incyte ID No 3143411CB1 36 gcggtccgtc ggtggcctag agatgctgct gccgcggttg cagttgtcgc gcacgcctct 60 gcccgccagc ccgctccacc gccgtagcgc ccgagtgtcg gggggcgcac ccgagtcggg 120 ccatgaggcc gggaaccgcg ctacaggccg tgctgctggc cgtgctgctg gtggggctgc 180 gggccgcgac gggtcgcctg ctgagtgggc agccagtctg ccggggaggg acacagaggc 240 cttgttataa agtcatttac ttccatgata cttctcgaag actgaacttt gaggaagcca 300 aagaagcctg caggagggat ggaggccagc tagtcagcat cgagtctgaa gatgaacaga 360 aactgataga aaagttcatt gaaaacctct tgccatctga tggtgacttc tggattgggc 420 tcaggaggcg tgaggagaaa caaagcaata gcacagcctg ccaggacctt tatgcttgga 480 ctgatggcag catatcacaa tttaggaact ggtatgtgga tgagccgtcc tgcggcagcg 540 aggtctgcgt ggtcatgtac catcagccat cggcacccgc tggcatcgga ggcccctaca 600 tgttccagtg gaatgatgac cggtgcaaca tgaagaacaa tttcatttgc aaatattctg 660 atgagaaacc agcagttcct tctagagaag ctgaaggtga ggaaacagag ctgacaacac 720 ctgtacttcc agaagaaaca caggaagaag atgccaaaaa aacatttaaa gaaagtagag 780 aagctgcctt gaatctggcc tacatcctaa tccccagcat tccccttctc ctcctccttg 840 tggtcaccac agttgtatgt tgggtttgga tctgtagaaa aagaaaacgg gagcagccag 900 accctagcac aaagaagcaa cacaccatct ggccctctcc tcaccaggga aacagcccgg 960 acctagaggt ctacaatgtc ataagaaaac aaagcgaagc tgacttagct gagacccggc 1020 cagacctgaa gaatatttca ttccgagtgt gttcgggaga agccactccc gatgacatgt 1080 cttgtgacta tgacaacatg gctgtgaacc catcagaaag tgggtttgtg actctggtga 1140 gcgtggagag tggatttgtg accaatgaca tttatgagtt ctccccagac caaatgggga 1200 ggagtaagga gtctggatgg gtggaaaatg aaatatatgg ttattaggac atataaaaaa 1260 ctgaaactga caacaatgga aaagaaatga taagcaaaat cctcttattt tctataagga 1320 aaatacacag aaggtctatg aacaagctta gatcaggtcc tgtggatgag catgtggtcc 1380 ccacgacctc ctgttggacc cccacgtttt ggctgtatcc tttatcccag ccagtcatcc 1440 agctcgacct tatgagaagg taccttgccc aggtctggca catagtagag tctcaataaa 1500 tgtcacttgg ttggttgtat ctaactttta agggacagag ctttacctgg cagtgataaa 1560 gatgggctgt ggagcttgga aaaccacctc tgttttcctt gctctataca gcagcacata 1620 ttatcataca gacagaaaat ccagaatctt ttcaaagccc acatatggta gcacaggctg 1680 gcctgtgcat cggcaattct catatctgtt tttttcaaag aataaaatca aataaagagc 1740 aggaaacaga aaaaaaaagg agatagggta gtagatgatg gtaaggagtg cttaagaagg 1800 tgtgggaaga gttgattgaa tggagggggt cccttgttt 1839 37 503 DNA Homo sapiens misc_feature Incyte ID No 3170835CB1 37 cggctcgaga tgaaattcgc catcgtcctg ttcgccctct ttgccgtggc cctggctgcc 60 cctactgtcg aggtcctgcg atcggatagc aatgttggaa tcgataacta ctcatatgca 120 gttgaaacca gcgacggtac ctcgaagagc gaggagggtg tgctgaagaa cgccggcacc 180 gagctagagg ccatctcaac ccacggctcc ttcagctacg tgggccctga tggccagacc 240 tacaccgtca cctacgtggc cgatgagaac ggattccagc cccagggtgc tcatctgccc 300 gttgcccccg ttgcctaagc taaggagttg aaataaattg tttataaaag caaaaaaact 360 ggcagaaaga acccaaagta cgtagacctt catgctagag agaatgtgat gtctgccgaa 420 gacaaccgta cccgcctgcg caatcttacg gccactagtg gatcctgcgc atgaggacac 480 cgagagcgag cgtggggggg ggc 503 38 2154 DNA Homo sapiens misc_feature Incyte ID No 3550808CB1 38 gaggacagga tgaggcccgg cctctcattt ctcctagccc ttctgttctt ccttggccaa 60 gctgcagggg atttggggga tgtgggacct ccaattccca gccccggctt cagctctttc 120 ccaggtgttg actccagctc cagcttcagc tccagctcca ggtcgggctc cagctccagc 180 cgcagcttag gcagcggagg ttctgtgtcc cagttgtttt ccaatttcac cggctccgtg 240 gatgaccgtg ggacctgcca gtgctctgtt tccctgccag acaccacctt tcccgtggac 300 agagtggaac gcttggaatt cacagctcat gttctttctc agaagtttga gaaagaactt 360 tctaaagtga gggaatatgt ccaattaatt agtgtgtatg aaaagaaact gttaaaccta 420 actgtccgaa ttgacatcat ggagaaggat accatttctt acactgaact ggacttcgag 480 ctgatcaagg tagaagtgaa ggagatggaa aaactggtca tacagctgaa ggagagtttt 540 ggtggaagct cagaaattgt tgaccagctg gaggtggaga taagaaatat gactctcttg 600 gtagagaagc ttgagacact agacaaaaac aatgtccttg ccattcgccg agaaatcgtg 660 gctctgaaga ccaagctgaa agagtgtgag gcctctaaag atcaaaacac ccctgtcgtc 720 caccctcctc ccactccagg gagctgtggt catggtggtg tggtgaacat cagcaaaccg 780 tctgtggttc agctcaactg gagagggttt tcttatctat atggtgcttg gggtagggat 840 tactctcccc agcatccaaa caaaggactg tattgggtgg cgccattgaa tacagatggg 900 agactgttgg agtattatag actgtacaac acactggatg atttgctatt gtatataaat 960 gctcgagagt tgcggatcac ctatggccaa ggtagtggta cagcagttta caacaacaac 1020 atgtacgtca acatgtacaa caccgggaat attgccagag ttaacctgac caccaacacg 1080 attgctgtga ctcaaactct ccctaatgct gcctataata accgcttttc atatgctaat 1140 gttgcttggc aagatattga ctttgctgtg gatgagaatg gattgtgggt tatttattca 1200 actgaagcca gcactggtaa catggtgatt agtaaactca atgacaccac acttcaggtg 1260 ctaaacactt ggtataccaa gcagtataaa ccatctgctt ctaacgcctt catggtatgt 1320 ggggttctgt atgccacccg tactatgaac accagaacag aagagatttt ttactattat 1380 gacacaaaca cagggaaaga gggcaaacta gacattgtaa tgcataagat gcaggaaaaa 1440 gtgcagagca ttaactataa cccttttgac cagaaacttt atgtctataa cgatggttac 1500 cttctgaatt atgatctttc tgtcttgcag aagccccagt aagctgttta ggagttaggg 1560 tgaaagagaa aatgtttgtt gaaaaaatag tcttctccac ttacttagat atctgcaggg 1620 gtgtctaaaa gtgtgttcat tttgcagcaa tgtttaggtg catagttcta ccacactaga 1680 gatctaggac atttgtcttg atttggtgag ttctcttggg aatcatctgc ctcttcaggc 1740 gcattttgca ataaagtctg tctagggtgg gattgtcaga ggtctagggg cactgtgggc 1800 ctagtgaagc ctactgtgag gaggcttcac tagaagcctt aaattaggaa ttaaggaact 1860 taaaactcag tatggcgtct agggattctt tgtacaggaa atattgccca atgactagtc 1920 cccatccatg tagcaccact aattcttcca tgcctggaag aaacctgggg acttagttag 1980 gtagattaat atctggagct cctcgaggga ccaaatctcc aacttttttt tcccctcact 2040 agcacctgga atgatgcttt gtatgtggca gataagtaaa tttggcatgc ttatatattc 2100 tacatcctga aagtggcctg gttttatgga gaagaggcct ctttatgcag tcaa 2154 39 733 DNA Homo sapiens misc_feature Incyte ID No 3683905CB1 39 gcgagcgagt tgccgagcgc gccccgtccc tcgcgcgcga tgctcccctg gacggcgctc 60 ggcctggccc tgagcttgcg gctggcgctg gcgcggagcg gcgcgggagc gcggtccacc 120 agcatcagcc ccccgagggg acctgatgtt cctgctggac agctcagcca gcgtctctca 180 ctacgagttc tcccgggttc gggagtttgt ggggcagctg gtggctccac tgcccctggg 240 caccggggcc ctgcgtgcca gtctggtgca cgtgggcagt cggccataca ccgagttccc 300 cttcggccag cacagctcgg gtgaggctgc ccaggatgcg gtgcgtgctt ctgcccagcg 360 catgggtgac acccacactg gcctggcgct ggtctatgcc aaggaacagc tgtttgctga 420 agcatcaggt gcccggccag gggtgcccaa agtgctggtg tgggtgacag atggcggctc 480 cagcgaccct gtgggccccc ccatgcagga gctcaaggac ctgggcgtca ccgtgttcat 540 tgtcagcacc ggccgaggca acttcctgga gctgtcagcc gctgcctcag cccctgccga 600 gaagcacctg cactttgtgg acgtggatga cctgcacatc attgtccaag agctgagggg 660 ctccattctc gacgcgatgc ggccgcaagc ttattccctt tagtgagggt taattttagc 720 ttgcactggc cgt 733 40 718 DNA Homo sapiens misc_feature Incyte ID No 4062841CB1 40 gccggcgcca gggcaggcgg gcggctggca gctgtggcgc cgacatggct gcgctggtgg 60 agccgctggg gctggagcgg gacgtgtccc gggcggttga gctcctcgag cggctccagc 120 gcagcgggga gctgccgccg cagaagctgc aggccctcca gcgagttctg cagagccgct 180 tctgctccgc tatccgagag gtgtatgagc agctttatga cacgctggac atcaccggca 240 gcgccgagat ccgagcccat gccacagcca aggccacagt ggctgccttc acagccagcg 300 agggccacgc acatcccagg gtagtggagc tacccaagac ggatgagggc ctaggcttca 360 acatcatggg tggcaaagag caaaactcgc ccatctacat ctcccgggtc atcccagggg 420 gtgtggctga ccgccatgga ggcctcaagc gtggggatca actgttgtcg gtgaacggtg 480 tgagcgttga gggtgagcag catgagaagg cggtggagct gctgaaggcg gcccagggct 540 cggtgaagct ggttgtccgt tacacaccgc gagtgctgga ggagatggag gcccggttcg 600 agaagatgcg ctctgcccgc cggcgccaac agcatcagag ctactcgtcc ttggagtctc 660 gaggttgaaa ccacagatct ggacgttcac gtgcactctc ttcctgtaca gtatttat 718 41 1235 DNA Homo sapiens misc_feature Incyte ID No 6394358CB1 41 gccgccggag ccgccgccgc agcggggacg gggagccccc gggggccccg ccaccgccgc 60 cgtccgccgt cacctacccg gactggatcg gccagagtta ctccgaggtg atgagcctca 120 acgagcactc catgcaggcg ctgtcctggc gcaagctcta cttgagccgc gccaagctta 180 aagcctccag ccggacctcg gctctgctct ccggcttcgc catggtggca atggtggagg 240 tgcagctgga cgctgaccac gactacccac cggggctgct catcgccttc agtgcctgca 300 ccacagtgct ggtggctgtg cacctgtttg cgctcatgat cagcacctgc atcctgccca 360 acatcgaggc ggtgagcaac gtgcacaatc tcaactcggt caaggagtcc ccccatgagc 420 gcatgcaccg ccacatcgag ctggcctggg ccttctccac cgtcatcggc acgctgctct 480 tcctagctga ggtggtgctg ctctgctggg tcaagttctt gcccctcaag aagcagccag 540 gccagccaag gcccaccagc aagccccccg ccagtggcgc agcagccaac gtcagcacca 600 gcggcatcac cccgggccag gcagctgcca tcgcctcgac caccatcatg gtgcccttcg 660 gcctgatctt tatcgtcttc gccgtccact tctaccgctc actggttagc cataagactg 720 accgacagtt ccaggagctc aacgagctgg cggagtttgc ccgcttacag gaccagctgg 780 accacagagg ggaccacccc ctgacgcccg gcagccacta tgcctaggcc catgtggtct 840 gggcccttcc agtgctttgg ccttacgccc ttccccttga ccttgtcctg ccccagcctc 900 acggacagcc tgcgcagggg gctgggcttc agcaaggggc agagcgtgga gggaagagga 960 tttttataag agaaatttct gcactttgaa actgtcctct aagagaataa gcatttcctg 1020 ttcttccagc tccaggtcca cctcctgttg ggaggcggtg gggggccaaa gtggggccac 1080 acactcgctg tgtcccctct cctcccctgt gccagtgcca cctgggtgcc tcctcctgtc 1140 ctgtccgtct caacctccct cccgtccagc attgagtgtg tacatgtgtg tgtgacacat 1200 aaatatactc ataaggacac ctccaaaaaa aaaaa 1235 42 11648 DNA Homo sapiens misc_feature Incyte ID No 2847752CB1 42 atggcgaggc ggccgccgtg gcggggcctc gggggacggt cgacccccat actcctgctc 60 cttctcctct ctttgttccc cctcagccag gaggagctgg ggggcggtgg gcaccagggc 120 tgggacccag gcttagctgc cactacgggg ccaagggcgc atatcggtgg cggagcctta 180 gctctttgtc cggagtcttc cggggtccgg gaggatgggg ggcctggcct gggggtcagg 240 gagcctatct tcgtggggct ccgagggaga aggcaaagcg cccggaatag tcgagggccc 300 cctgagcagc cgaatgagga gctggggatt gaacacggcg tccagccatt gggcagccgc 360 gaacgagaga caggacaggg accagggtct gtgttatact ggcgcccaga ggtctcctct 420 tgcgggcgga caggaccttt gcaaagaggt agtctgtcac caggggctct gtcctcaggg 480 gtcccgggct cggggaacag ctcgcccctc ccttcagact ttttgattcg gcaccacggt 540 cccaagccgg tgtcctccca gcggaacgct gggacaggct cccgcaaaag agtgggcacc 600 gcgcgctgct gtggggaatt atgggcaaca gggagcaagg gtcagggcga gagagccacg 660 acatccggag cagaaaggac agccccccgg cggaactgtc ttccaggggc ctcgggatct 720 ggccccgagc tggattcagc accacgcacg gcgaggacag ctcctgcatc aggttcagca 780 ccccgcgagt ctcggacagc tcccgagccg gcgcccaagc gcatgcgctc ccggggtctc 840 ttccgctgcc gcttcctccc gcagcgcccc gggccgcgtc ccccgggact cccggcccgt 900 cctgaagcca ggaaagtaac ctcggcgaac cgggcacgct ttcgtcgcgc cgcaaaccgc 960 cacccgcagt ttccgcagta caactaccag acgctggtgc cggagaatga ggcagcaggc 1020 accgcggtgc tacgcgtggt tgctcaggac ccggacgccg gcgaggccgg gcgcctagtc 1080 tactcgctgg cggcactcat gaacagccgc tcgctggagc tgttcagcat cgacccgcag 1140 agcggcctta tccgtacggc ggcagctctg gaccgcgaga gcatggagcg tcactacctg 1200 cgtgtgaccg cgcaggacca cgggtcgccg cgcctctcgg ccaccacgat ggtggccgtg 1260 acagtagccg accgcaacga ccactcgccg gtttttgagc aagcgcagta ccgggagacc 1320 cttcgcgaga atgtggagga gggctaccct atcctgcagc tgcgtgccac tgacggcgac 1380 gcgcccccca acgccaacct gcgctaccgc ttcgtggggc cgccagctgc gcgcgctgca 1440 gctgccgccg ccttcgagat tgatccacgc tccggcctca tcagcaccag cggccgagtg 1500 gaccgcgagc acatggaaag ctatgagctg gtggtggaag ccagcgacca gggccaggaa 1560 cccgggccgc gctcggccac tgtgcgcgta cacataactg tgctagacga gaacgacaat 1620 gctcctcagt tcagcgagaa gcgctacgtg gcgcaggtgc gcgaggatgt gcgcccccac 1680 acagtcgtgc tgcgcgtcac ggccactgac cgggacaagg acgccaacgg attggtgcac 1740 tacaacatca tcagtggcaa tagccgtgga cactttgcca tcgacagcct cactggcgag 1800 atccaggtgg tggcacctct ggacttcgag gcagagagag agtatgcctt gcgcatcagg 1860 gcgcaggatg ctggccggcc accgctgtcc aacaacacgg gcctggccag catccaggtg 1920 gtggacatca atgaccacat tcctattttt gtcagcacgc ccttccaagt ttctgtcttg 1980 gaaaatgctc ccttgggtca ctcagtcatc cacattcagg cagtcgatgc agaccatggg 2040 gagaatgcca gattggagta ctccctaact ggtgtggcac ctgatactcc ttttgtgata 2100 aacagcgcca ctggctgggt ctctgtgagt ggtcccctgg accgtgagtc tgtggagcat 2160 tacttctttg gtgtggaggc tcgagaccat ggctcacccc cactctctgc ctcagccagt 2220 gtcaccgtga ctgtgctgga cgttaatgac aatcggcctg agttcacaat gaaggagtac 2280 cacctacgac tgaatgagga tgcagctgtg ggcaccagtg tggtcagcgt gaccgcagta 2340 gaccgtgatg ccaacagtgc catcagctac cagatcacag gcggcaacac ccggaatcgc 2400 tttgccatca gcacccaggg gggtgtgggt ctggtgactc tggctctgcc actggactac 2460 aagcaggaac gctacttcaa gctggtacta actgcatctg accgtgccct tcatgatcac 2520 tgctatgtgc acatcaacat cacagatgcc aacactcatc ggccggtctt tcaaagtgcc 2580 cactactcag tgagtgtgaa tgaagatcgg ccaatgggta gcaccatagt ggtcatcagt 2640 gcctctgatg atgacgtggg tgagaatgct cgtatcacct atctcctgga ggacaacctg 2700 ccccagttcc gcattgatgc agactcagga gccattacat tacaggcccc attagactat 2760 gaggaccagg tgacctacac cctggctatc acagctcggg acaatggcat cccacagaag 2820 gcagacacta cttatgtgga ggtgatggtc aatgacgtga atgacaatgc tccacaattt 2880 gtggcctccc actatacagg gctggtctct gaggatgccc cacctttcac cagtgtcctg 2940 cagatctcag ccactgaccg ggatgctcat gccaatggcc gggtccagta cactttccag 3000 aatggtgaag atggggatgg agattttacc attgagccca cctctggaat tgtccgtaca 3060 gtaaggcggc tagaccggga ggcagtatca gtgtatgagt tgactgccta cgcagtggac 3120 agaggtgtgc ccccactccg gactccagtc agtatccagg tgatggtgca ggatgtgaac 3180 gacaatgcac ctgtcttccc agctgaggag tttgaggtgc gggtgaaaga gaatagcatt 3240 gtgggctcag tggtggccca gatcactgca gtggaccctg acgaaggccc caatgcccat 3300 ataatgtacc agatcgtgga ggggaacatc cctgagctgt tccaaatgga catcttctct 3360 ggagaactga cggcactcat tgacctagac tatgaggctc gccaagaata tgtgattgtg 3420 gtgcaggcca catctgctcc tttggtcagc cgggccactg tgcacgtccg cctggttgac 3480 cagaatgaca acagccctgt gctcaacaac ttccagatcc tcttcaacaa ctatgtatcc 3540 aaccgttcag acaccttccc gtcgggcatt attgggcgca tcccagctta tgaccccgat 3600 gtctccgacc acctcttcta ctcctttgag cgtggcaatg agctgcagct gctggtagtc 3660 aaccagacca gtggggagct gcgactcagc cgaaagctag acaataaccg cccactggtg 3720 gcctccatgt tggtgactgt cacagatggc ctgcacagcg tgacggcgca gtgtgtgctg 3780 cgcgtggtca tcatcacgga ggagttgctg gccaacagcc tgaccgtgcg ccttgagaac 3840 atgtggcagg agcgcttcct gtcaccgctg ctgggccgct tcctcgaggg cgtggctgcg 3900 gtgctcgcta cgcccgctga ggacgtcttc atcttcaaca tccagaacga cacagacgta 3960 gggggcaccg tgctcaatgt gagtttctcg gcgctagctc cacgtggggc cggggcgggc 4020 gctgcagggc cctggttcag ctccgaggag ctgcaagagc agttgtacgt gcgccgggcg 4080 gcgctggcgg ctcgctccct gctcgacgta ctgcccttcg acgacaacgt gtgcctgcga 4140 gagccctgtg agaactacat gaaatgcgtg tccgtgctcc gctttgactc gtccgcgccc 4200 ttcctggcct cggcctccac gctgttccga cccatccagc ccatcgctgg cctgcgctgc 4260 cgctgcccgc ccggattcac gggagacttt tgcgagaccg agctcgacct ctgctactcc 4320 aacccatgtc gcaacggcgg agcctgcgcg cggcgcgagg gaggctacac gtgcgtctgc 4380 cgcccgcgct tcaccggaga ggactgcgag ctggacaccg aggccggccg ctgcgtgccg 4440 ggcgtctgcc gcaacggggg cacctgcacc gacgcgccca acggcggctt tcgctgccag 4500 tgcccggcag gcggcgcctt cgagggcccg cgctgcgagg tggctgcgcg ctccttcccg 4560 cccagttcgt tcgtcatgtt tcgcggcctg cggcagcgat tccaccttac gctgtccctc 4620 tcgttcgcga cagtgcagca gagcgggctg ctcttctaca acgggcgcct gaacgagaag 4680 cacgacttcc tggccctgga actcgtggct ggccaagtgc ggctcacata ttccacgggt 4740 gaatccaaca ccgtggtcag ccccacagtt ccagggggct tgagtgacgg gcaatggcat 4800 acagtgcatc tgagatacta caacaagccc cggacagatg ccctaggggg tgcacagggc 4860 ccctccaagg acaaggtggc tgtgctaagc gtggatgatt gtgatgtggc cgtggctctg 4920 cagtttggtg ctgagattgg caactactca tgcgcggctg ctggtgtgca aacaagctcc 4980 aagaagtccc tggacctgac gggccctctt cttctgggag gtgtccccaa cctccccgag 5040 aacttccccg tatcccataa ggacttcatc ggctgtatgc gggacctgca cattgatggc 5100 cgccgagtgg acatggcggc ttttgtcgca aataatggca ccatggcagg ctgccaagcc 5160 aagctacact tttgtgactc aggcccctgc aagaacagtg gcttctgctc ggagcgctgg 5220 ggcagcttca gctgcgactg ccctgtgggc ttcggcggca aagactgtca gcttactatg 5280 gcccatcccc accatttccg tggcaacggc acactgagct ggaactttgg aagtgacatg 5340 gctgtgtctg tgccatggta cctggggctg gcatttcgga cacgggcaac gcagggggtc 5400 ctgatgcaag tgcaggctgg gccacacagc acgctccttt gccagctaga tcgggggtta 5460 ctgtctgtga cagtgaccag gggctcgggc cgtgcttccc atctccttct ggaccaggtg 5520 actgtcagtg atggccggtg gcacgatctg cggctggagt tgcaggagga accaggtggc 5580 cggcggggcc accatgtcct tatggtctca ctggacttta gcctcttcca ggacaccatg 5640 gcggtgggga gtgagctgca gggcctgaag gtaaagcagc tccacgtggg aggcctgccc 5700 cccggcagtg cagaggaggc tcctcagggt ctggttggct gcatccagcc accgagtgaa 5760 tgcggacctg gctgtgttgt gaccaacgcc tgtgcctctg ggccctgccc acctcacgca 5820 gactgccggg acctctggca gaccttttct tgcacctgcc agccaggtta ctacggccca 5880 ggctgtgtgg atgcctgcct cctgaacccc tgtcagaacc agggatcatg ccggcacctg 5940 ccaggagccc cccatggcta tacctgtgac tgtgtgggtg gctatttcgg gcaccactgt 6000 gagcacagga tggaccagca gtgcccacgg ggctggtggg ggagcccaac ctgtggcccc 6060 tgcaactgtg atgttcacaa aggttttgat cccaactgca acaagacaaa tgggcagtgt 6120 cactgcaagg agttccacta ccgaccgcgg ggcagtgact cttgcctccc atgtgactgc 6180 taccctgtgg gctccacctc gcgctcatgt gcaccccaca gcgggcagtg cccctgtcgc 6240 ccaggagccc ttggccgcca gtgcaacagc tgtgacagtc ccttcgcaga ggtgacagcc 6300 agcggctgcc gggtgctcta tgatgcctgc cctaagtccc tgagatctgg tgtgtggtgg 6360 ccccagacaa agtttggcgt cctggccaca gtgccctgtc cccggggggc cctgggattg 6420 cggggtgcag gtgctgctgt gcggctgtgt gatgaggccc agggttggct ggagcccgac 6480 ctcttcaact gtacctcccc tgcctttcga gagctcagtc tgctgctgga tggcctagag 6540 ctgaacaaga cggcactgga taccatggag gccaagaagc tggctcagcg gctacgggag 6600 gtgactggcc acactgacca ctattttagc caagatgttc gagtcactgc ccgcctgctg 6660 gcccacctgc tggccttcga gagccatcag cagggcttcg ggctgacagc cacacaggat 6720 gcccacttca atgagaatct gctgtgggcc ggctctgcac tgcttgcccc agagacaggg 6780 gacttgtggg cggcgctggg gcagcgggcc cctgggggct ccccaggcag cgcgggactg 6840 gtgaggcacc tggaggagta tgcagccaca ctcgcaagga atatggaact cacatacctg 6900 aatcccatgg ggctggtgac gcctaatatc atgctcagca ttgaccgcat ggagcacccc 6960 agttctcccc ggggggcccg tcgctaccct cgctaccata gcaacctctt tcgaggccag 7020 gatgcctggg atcctcacac ccatgtgctg ctgccttccc agtccccacg gccatcccca 7080 tctgaagttc tgcccacaag cagcagcata gaaaactcca ccacctcaag tgtggtcccc 7140 ccaccagccc cgccagagcc agagcctggg atctccatta tcattctcct cgtttaccgc 7200 accttagggg gactgctccc tgcccagttc caggcagaac gccgaggtgc caggcttcct 7260 cagaaccccg tcatgaactc cccggtggtc agcgtggctg tgttccacgg acgcaacttc 7320 ctaaggggaa tcctggagtc ccccatcagc ctagagtttc gcctgctaca gacagcgaat 7380 cggagcaagg cgatctgtgt gcagtgggac ccacctggcc tggcggagca gcatggtgtg 7440 tggacagcac gggactgcga gctggtgcac aggaatgggt cccacgcacg gtgtcgctgc 7500 agccggacag ggacctttgg ggtcctcatg gatgcctctc cccgtgagag gctggagggc 7560 gacctggagc tgctggctgt gttcacccac gtggtcgtgg ctgtgtctgt ggctgcgctg 7620 gtgctgactg cagccatcct gctgagcctg cgcagcctca agtccaatgt gcgtgggatc 7680 catgccaatg tggcagccgc cctgggggtg gcagagctcc tcttcctgct ggggattcac 7740 aggacccaca atcagctggt gtgcactgca gtcgccatcc tcctgcacta cttcttcctc 7800 agcaccttcg cgtggctctt cgtgcagggg ctgcacctct accgcatgca ggttgagcca 7860 cgcaacgtgg accgcggcgc catgcgcttc taccatgccc tgggctgggg cgtccctgct 7920 gtgctgctgg gccttgctgt gggcctggac cctgagggct atgggaaccc tgacttctgc 7980 tggatctcag tccacgagcc cctcatctgg agctttgctg gccctgttgt cctggtcata 8040 gtgatgaacg ggaccatgtt tctcctcgct gcccgcacat cctgctccac agggcagagg 8100 gaggccaaga agacctctgc actgaccctt cgcagctcct tcctgctgct tctgctggtc 8160 agtgcctcct ggctctttgg gctcctggca gtcaaccaca gcatcctagc cttccactac 8220 ctccatgctg gactctgcgg cctccagggc ctggcggtgc tgctgctctt ctgtgtccta 8280 aatgcagatg ctcgggctgc ctggatgcca gcctgtctgg gcaggaaggc agcgcctgag 8340 gaggcaaggc cagcacctgg gctgggacct ggggcctaca acaacacggc tctctttgag 8400 gagagtggcc tcatccgcat cactctgggc gcctccaccg tctcctctgt gagcagtgcc 8460 cgctccggcc ggacccagga ccaggacagc cagcggggcc gcagctacct cagggacaat 8520 gtcctggttc gacatggctc agccgctgac cacactgacc acagcctcca ggctcatgct 8580 ggccccactg acctggacgt ggccatgttc catcgagatg ctggcgcaga ctccgactct 8640 gacagtgacc tgtccttgga ggaggagagg agtctctcca ttccatcttc agaaagcgag 8700 gacaatggcc ggacgcgggg gcgcgtccaa cggccactct gccgagcagc ccagagtgag 8760 aggctcctca cccaccccaa agatgtggat ggcaatgacc tcctgtccta ctggccagcc 8820 ctgggggagt gcgaggcagc cccctgtgct ctgcagactt ggggctctga aaggcgcctg 8880 gggctggaca ccagcaagga tgcagctaac aacaaccagc cagacccggc cctgaccagt 8940 ggggatgaga cttctctggg ccgggcccag cgccagagga aaggcatcct gaagaaccgg 9000 ttgcaatacc cactggtgcc acagacccga ggtgcccctg agctgtcctg gtgccgtgca 9060 gccaccttgg gccaccgtgc tgtgccagct gcctcttacg gtcgcatcta tgctggcggg 9120 ggcacgggca gcctttcaca gccagccagc cgctactctt ctagagaaca gctggacctg 9180 ctcctccggc ggcaactgag ccgtgagcga ctagaggaag cccctgcccc tgttctacgt 9240 cccctgagcc ggccagggtc ccaggaatgc atggatgctg caccaggccg actggagccc 9300 aaagatcggg gcagcaccct gccacggagg cagccacctc gggactaccc tggcgccatg 9360 gctggccgct tcgggtcacg ggatgcgctc gacttagggg cacctcgaga gtggttgagc 9420 acgctgcctc cgccccgccg cacccgggac cttgacccac agcccccacc tctgcccctg 9480 tctccccagc ggcaactctc aagggacccc ctcttgccat cccggccgct ggactctctg 9540 tctaggagct cgaactctcg ggagcagctg gaccaggtgc ctagccggca cccctcacga 9600 gaagcccttg ggccactccc gcagctgctc agagctaggg aggactcggt cagtggcccc 9660 agccatggcc cctccacaga acagttggac attctttcct ccatccttgc ctctttcaac 9720 tcctcggccc tctcctctgt gcaatcttca agcacaccct tgggccctca caccactgcc 9780 acaccttctg ccacagcctc tgtgcttggg ccctccacgc cacgttctgc cacgtctcac 9840 agcatctcgg agctgtcgcc agactcagaa gttcccagaa gtgagggtca ctcctgaggg 9900 gatgacggcg tggacgagga acagctgagg gcgacagagg atctaggcta acaggagaga 9960 ctccaggagt gggggcagat cccaaggcag cctcctgctc cccagtggtg ggtgccccag 10020 ctctacctgg tgtggcaggg ctgaggctcc atgtgcatct gtgagcatgc gtgtgacagg 10080 tgcagagacg ggggactgga gggagacttt tatacgtttt gtacctttgt aaccagagag 10140 atgcttatgt tatttttcag cttttctgtc tcctgggggg tttgaggctg ggctgggagg 10200 gggagggaga tagagggaga gatgcagttt gaccccattt gggtcctgag caaaccctat 10260 gctcatctct ctctccttcc tggggtggac tcagatgggt gggacacatg ccttcctccc 10320 cctattccac ccccaagttg atctgagtat cgtcaggggc ccaaagtaca gaattgttct 10380 ttgcttttta ttgaatgctc caaaggccaa acttctgggg ctgggggttg gtcttggaaa 10440 caggggtcct ctgacttcct catgggggct tgctcatacc gcccctcctg gtggatgtgt 10500 gtgtttatta tgtggagtcc ctgccactta ctgccttatg acctaggact gatgctgtgg 10560 ggtgctggtg gagcagctga tgtcgtgttt acagagcaag gcttccctgt ctcccacggg 10620 gaggggctcg ggcctctagt cagacattcc tgcagagggt cggtggaggg gtcattcacc 10680 tgcccctgca gcaagcaaaa gttgtctgtg gtgccatttg attccctgac actgccccct 10740 gcttgaattg attccgaagg gtagggtggg aaggtgagca aagggagcag aaacaaggga 10800 attcaagacc cagaatgtag gtgccactgc ctcctatgtt tacaggatcc tccgtggccc 10860 taggcacctg ggctgcagga agtgactccg ttccactcct cctttattcc cttaaaaagg 10920 gaaaaatgac tgttacgacc ctgttcacaa aactcttact tttgctattt tgtctgctgt 10980 ccagaactga agactttaaa attttgttac tgtttacaag tccagattca aaaaatgttt 11040 ttactttgtt tacaactcaa aactttgagt tttacacttt gtttacagta gataattttt 11100 tttcctttgt ttccaagtga aaggtaggga aagtgggaga gggacttgga ggacccacct 11160 gtgaggaccc tgacctggcc atcttgaggg gttttctaac ccccaggtct cccaggccga 11220 aggtcagcct tgagtcccgt ttaacagcag atccagaaga ccttgagagt aggcgtcctc 11280 taaccacggg ggagagtggc tgtgcagggc tggggggtgg tctgtgcaga cacctcctca 11340 cccaccaccc catgcatact cttgggaagc agcttcctgg gagattagaa attctacttc 11400 cctgactgga gctaaatccc accagccagg acccaaactc tccttaccga gaaggacccc 11460 agctcttgaa gggctgagtg gcctgctggg ggtgggaggg tgtctttact atgtcctagg 11520 tttcgtagat gcccctctct ggggttcccc tcctccagcc cagcggccct ctttcctgtc 11580 tgtgtaaatt gttccgtgaa gccgcgctct gttttgggaa taaacttcta tagaaaacaa 11640 aaaaaaaa 11648 

What is claimed is:
 1. An isolated polypeptide comprising an amino acid sequence selected from the group consisting of: a) an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-21.
 2. An isolated polypeptide of claim 1 selected from the group consisting of SEQ ID NO:1-21.
 3. An isolated polynucleotide encoding a polypeptide of claim
 1. 4. An isolated polynucleotide encoding a polypeptide of claim
 2. 5. An isolated polynucleotide of claim 4 selected from the group consisting of SEQ ID NO:22-42.
 6. A recombinant polynucleotide comprising a promoter sequence operably linked to a polynucleotide of claim
 3. 7. A cell transformed with a recombinant polynucleotide of claim
 6. 8. A transgenic organism comprising a recombinant polynucleotide of claim
 6. 9. A method for producing a polypeptide of claim 1, the method comprising: a) culturing a cell under conditions suitable for expression of the polypeptide, wherein said cell is transformed with a recombinant polynucleotide, and said recombinant polynucleotide comprises a promoter sequence operably linked to a polynucleotide encoding the polypeptide of claim 1, and b) recovering the polypeptide so expressed.
 10. An isolated antibody which specifically binds to a polypeptide of claim
 1. 11. An isolated polynucleotide comprising a polynucleotide sequence selected from the group consisting of: a) a polynucleotide sequence selected from the group consisting of SEQ ID NO:22-42, b) a naturally occurring polynucleotide sequence having at least 90% sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID NO:22-42, c) a polynucleotide sequence complementary to a), d) a polynucleotide sequence complementary to b), and e) an RNA equivalent of a)-d).
 12. An isolated polynucleotide comprising at least 60 contiguous nucleotides of a polynucleotide of claim
 11. 13. A method for detecting a target polynucleotide in a sample, said target polynucleotide having a sequence of a polynucleotide of claim 11, the method comprising: a) hybridizing the sample with a probe comprising at least 20 contiguous nucleotides comprising a sequence complementary to said target polynucleotide in the sample, and which probe specifically hybridizes to said target polynucleotide, under conditions whereby a hybridization complex is formed between said probe and said target polynucleotide or fragments thereof, and b) detecting the presence or absence of said hybridization complex, and, optionally, if present, the amount thereof.
 14. A method of claim 13, wherein the probe comprises at least 60 contiguous nucleotides.
 15. A method for detecting a target polynucleotide in a sample, said target polynucleotide having a sequence of a polynucleotide of claim 11, the method comprising: a) amplifying said target polynucleotide or fragment thereof using polymerase chain reaction amplification, and b) detecting the presence or absence of said amplified target polynucleotide or fragment thereof, and, optionally, if present, the amount thereof.
 16. A composition comprising an effective amount of a polypeptide of claim 1 and a pharmaceutically acceptable excipient.
 17. A composition of claim 16, wherein the polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1-21.
 18. A method for treating a disease or condition associated with decreased expression of functional XMAD, comprising administering to a patient in need of such treatment the composition of claim
 16. 19. A method for screening a compound for effectiveness as an agonist of a polypeptide of claim 1, the method comprising: a) exposing a sample comprising a polypeptide of claim 1 to a compound, and b) detecting agonist activity in the sample.
 20. A composition comprising an agonist compound identified by a method of claim 19 and a pharmaceutically acceptable excipient.
 21. A method for treating a disease or condition associated with decreased expression of functional XMAD, comprising administering to a patient in need of such treatment a composition of claim
 20. 22. A method for screening a compound for effectiveness as an antagonist of a polypeptide of claim 1, the method comprising: a) exposing a sample comprising a polypeptide of claim 1 to a compound, and b) detecting antagonist activity in the sample.
 23. A composition comprising an antagonist compound identified by a method of claim 22 and a pharmaceutically acceptable excipient.
 24. A method for treating a disease or condition associated with overexpression of functional XMAD, comprising administering to a patient in need of such treatment a composition of claim
 23. 25. A method of screening for a compound that specifically binds to the polypeptide of claim 1, said method comprising the steps of: a) combining the polypeptide of claim 1 with at least one test compound under suitable conditions, and b) detecting binding of the polypeptide of claim 1 to the test compound, thereby identifying a compound that specifically binds to the polypeptide of claim
 1. 26. A method of screening for a compound that modulates the activity of the polypeptide of claim 1, said method comprising: a) combining the polypeptide of claim 1 with at least one test compound under conditions permissive for the activity of the polypeptide of claim 1, b) assessing the activity of the polypeptide of claim 1 in the presence of the test compound, and c) comparing the activity of the polypeptide of claim 1 in the presence of the test compound with the activity of the polypeptide of claim 1 in the absence of the test compound, wherein a change in the activity of the polypeptide of claim 1 in the presence of the test compound is indicative of a compound that modulates the activity of the polypeptide of claim
 1. 27. A method for screening a compound for effectiveness in altering expression of a target polynucleotide, wherein said target polynucleotide comprises a sequence of claim 5, the method comprising: a) exposing a sample comprising the target polynucleotide to a compound, under conditions suitable for the expression of the target polynucleotide, b) detecting altered expression of the target polynucleotide, and c) comparing the expression of the target polynucleotide in the presence of varying amounts of the compound and in the absence of the compound.
 28. A method for assessing toxicity of a test compound, said method comprising: a) treating a biological sample containing nucleic acids with the test compound; b) hybridizing the nucleic acids of the treated biological sample with a probe comprising at least 20 contiguous nucleotides of a polynucleotide of claim 11 under conditions whereby a specific hybridization complex is formed between said probe and a target polynucleotide in the biological sample, said target polynucleotide comprising a polynucleotide sequence of a polynucleotide of claim 11 or fragment thereof; c) quantifying the amount of hybridization complex; and d) comparing the amount of hybridization complex in the treated biological sample with the amount of hybridization complex in an untreated biological sample, wherein a difference in the amount of hybridization complex in the treated biological sample is indicative of toxicity of the test compound.
 29. A diagnostic test for a condition or disease associated with the expression of XMAD in a biological sample, the method comprising: a) combining the biological sample with an antibody of claim 10, under conditions suitable for the antibody to bind the polypeptide and form an antibody:polypeptide complex, and b) detecting the complex, wherein the presence of the complex correlates with the presence of the polypeptide in the biological sample.
 30. The antibody of claim 10, wherein the antibody is: a) a chimeric antibody, b) a single chain antibody, c) a Fab fragment, d) a F(ab′)₂ fragment, or e) a humanized antibody.
 31. A composition comprising an antibody of claim 10 and an acceptable excipient.
 32. A method of diagnosing a condition or disease associated with the expression of XMAD in a subject, comprising administering to said subject an effective amount of the composition of claim
 31. 33. A composition of claim 31, wherein the antibody is labeled.
 34. A method of diagnosing a condition or disease associated with the expression of XMAD in a subject, comprising administering to said subject an effective amount of the composition of claim
 33. 35. A method of preparing a polyclonal antibody with the specificity of the antibody of claim 10, the method comprising: a) immunizing an animal with a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, or an immunogenic fragment thereof, under conditions to elicit an antibody response, b) isolating antibodies from said animal, and c) screening the isolated antibodies with the polypeptide, thereby identifying a polyclonal antibody which binds specifically to a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-21.
 36. An antibody produced by a method of claim
 35. 37. A composition comprising the antibody of claim 36 and a suitable carrier.
 38. A method of making a monoclonal antibody with the specificity of the antibody of claim 10, the method comprising: a) immunizing an animal with a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-21, or an immunogenic fragment thereof, under conditions to elicit an antibody response, b) isolating antibody producing cells from the animal, c) fusing the antibody producing cells with immortalized cells to form monoclonal antibody-producing hybridoma cells, d) culturing the hybridoma cells, and e) isolating from the culture monoclonal antibody which binds specifically to a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-21.
 39. A monoclonal antibody produced by a method of claim
 38. 40. A composition comprising the antibody of claim 39 and a suitable carrier.
 41. The antibody of claim 10, wherein the antibody is produced by screening a Fab expression library.
 42. The antibody of claim 10, wherein the antibody is produced by screening a recombinant immunoglobulin library.
 43. A method of detecting a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-21 in a sample, the method comprising: a) incubating the antibody of claim 10 with a sample under conditions to allow specific binding of the antibody and the polypeptide, and b) detecting specific binding, wherein specific binding indicates the presence of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-21 in the sample.
 44. A method of purifying a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-21 from a sample, the method comprising: a) incubating the antibody of claim 10 with a sample under conditions to allow specific binding of the antibody and the polypeptide, and b) separating the antibody from the sample and obtaining the purified polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-21.
 45. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:1.
 46. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:2.
 47. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:3.
 48. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:4.
 49. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:5.
 50. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:6.
 51. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:7.
 52. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:8.
 53. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:9.
 54. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:10.
 55. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:11.
 56. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:12.
 57. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:13.
 58. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:14.
 59. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:15.
 60. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:16.
 61. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:17.
 62. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:18.
 63. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:19.
 64. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:20.
 65. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:21.
 66. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:22.
 67. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:23.
 68. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:24.
 69. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:25.
 70. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:26.
 71. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:27.
 72. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:28.
 73. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:29.
 74. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:30.
 75. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:31.
 76. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:32.
 77. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:33.
 78. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:34.
 79. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:35.
 80. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:36.
 81. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:37.
 82. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:38.
 83. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:39.
 84. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:40.
 85. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:41.
 86. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:42.
 87. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:1.
 88. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:2.
 89. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:3.
 90. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:4.
 91. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:5.
 92. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:6.
 93. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:7.
 94. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:8.
 95. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:9.
 96. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:10.
 97. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:11.
 98. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:12.
 99. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:13.
 100. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:14.
 101. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:15.
 102. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:16.
 103. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:17.
 104. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:18.
 105. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:19.
 106. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:20.
 107. A method of claim 9, wherein the polypeptide has the sequence of SEQ ID NO:21.
 108. A microarray wherein at least one element of the microarray is a polynucleotide of claim
 12. 109. A method for generating a transcript image of a sample which contains polynucleotides, the method comprising the steps of: a) labeling the polynucleotides of the sample, b) contacting the elements of the microarray of claim 108 with the labeled polynucleotides of the sample under conditions suitable for the formation of a hybridization complex, and c) quantifying the expression of the polynucleotides in the sample.
 110. An array comprising different nucleotide molecules affixed in distinct physical locations on a solid substrate, wherein at least one of said nucleotide molecules comprises a first oligonucleotide or polynucleotide sequence specifically hybridizable with at least 30 contiguous nucleotides of a target polynucleotide, said target polynucleotide having a sequence of claim
 11. 111. An array of claim 110, wherein said first oligonucleotide or polynucleotide sequence is completely complementary to at least 30 contiguous nucleotides of said target polynucleotide.
 112. An array of claim 110, wherein said first oligonucleotide or polynucleotide sequence is completely complementary to at least 60 contiguous nucleotides of said target polynucleotide.
 113. An array of claim 110, which is a microarray.
 114. An array of claim 110, further comprising said target polynucleotide hybridized to said first oligonucleotide or polynucleotide.
 115. An array of claim 110, wherein a linker joins at least one of said nucleotide molecules to said solid substrate.
 116. An array of claim 110, wherein each distinct physical location on the substrate contains multiple nucleotide molecules having the same sequence, and each distinct physical location on the substrate contains nucleotide molecules having a sequence which differs from the sequence of nucleotide molecules at another physical location on the substrate. 